Sulfur (vi) fluoride compounds and methods for the preparation thereof

ABSTRACT

This application describes modified amino acids and polypeptides comprising a SO 2 F or CH 2 CH 2 SO 2 F group bound to the side chain of an amono acid or amino acid residue of a polypeptide in place of a hydrogen of a hydroxyl or amino substituent thereof. Methods of covalently binding the polypeptides to receptot sites of receptor proteins are also described herein.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.17/012,783, filed on Sep. 4, 2020, which is a division of U.S.application Ser. No. 16/158,608, filed on Oct. 12, 2018, now U.S. Pat.No. 10,765,645, which is a division of U.S. application Ser. No.15/316,742, filed on Dec. 6, 2016, now U.S. Pat. No. 10,117,840, whichis a 371 of PCT/US2015/034516, filed Jun. 5, 2015, which claims prioritybenefit to U.S. Provisional Application No. 62/008,925, filed on Jun. 6,2014, each of which is incorporated herein by reference in theirentirety.

STATEMENT OF GOVERNMENT LICENSE RIGHTS

This invention was made with government support from National Institutesof Health, Grants No. U01NS058046 and EB015663, and National ScienceFoundation, Grant No. CHE 1011796. The government has certain rights inthis invention.

INCORPORATION OF SEQUENCE LISTING

Biological sequence information for this application is included in anASCII text file having the file name “TSRI-1613-1-SEQ-V2.TXT”, createdon Feb. 28, 2018, and having a file size of 4,924 bytes, which isincorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to sulfur(VI) fluoride compounds,including therapeutic compounds and compositions, as well as and methodsof using and producing the compounds and compositions.

BACKGROUND

“Click” chemistry was introduced as a conceptual framework forfunctional molecular assembly a decade ago, emphasizing the importanceof carbon-heteroatom linkages in joining modular building blocks (see H.C. Kolb, M. G. Finn, K. B. Sharpless, Angew. Chem. 2001, 123, 2056-2075;Angew. Chem. Int. Ed 2001, 2040, 2004-2021). Taking inspiration fromnature, click reactions were identified as processes that work underoperationally simple, oxygen- and water-friendly conditions, andgenerate products in high yields with minimal requirements for productpurification. Such reactions invariably have an unusual combination ofstrong thermodynamic driving forces and consistent, well-controlledreaction pathways. In tandem, these two features allow the use of widelyvarying substrates with great reliability.

The azide-alkyne cycloaddition reaction (see R. Huisgen, Angew. Chem.1963, 75, 604-637; Angew. Chem. Int. Ed Engl. 1963, 1962, 1565-1598) isespecially useful because of the unobtrusive nature of its participatingfunctional groups and the ability to turn on their ligating ability (todifferent extents, and for different purposes) by Cu(I) catalysts (see(a) C. W. Tornøe, C. Christensen, M. Meldal, J. Org. Chem. 2002, 67,3057-3062. (b) V. V. Rostovtsev, L. G. Green, V. V. Fokin, K. B.Sharpless, Angew. Chem. 2002, 114, 2708-2711; Angew. Chem. Int. Ed 2002,2741, 2596-2599.), installing strain in the alkyne component (see (a) G.Wittig, A. Krebs, Chem. Ber. Recl. 1961, 94, 3260-3275. (b) N. J. Agard,J. A. Prescher, C. R. Bertozzi, J. Am. Chem. Soc. 2004, 126,15046-15047), or holding them in close spatial proximity (see (a) W. G.Lewis, L. G. Green, F. Grynszpan, Z. Radic, R. P. Carlier, P. Taylor, M.G. Finn, K. B. Sharpless, Angew. Chem. 2002, 114, 1095-1099; Angew.Chem. Int. Ed 2002, 1041, 1053-1057. (b) H. D. Agnew, R. D. Rhode, S. W.Millward, A. Nag, W. S. Yeo, J. E. Hein, S. M. Pitram, A. A. Tariq, V.M. Burns, R. J. Krom, V. V. Fokin, K. B. Sharpless, J. R. Heith, Angew.Chem. 2009, 121, 5044-5048; Angew. Chem. Int. Ed 2009, 5048, 4944-4948).Thus, this click reaction emerged by finding ways to induce twofunctional groups to react with each other that otherwise have verylittle propensity to react with anything, in spite of their highlyenergetic nature. In contrast, most other click reactions find a usefulwindow of activity by moderating the properties of at least one highlyreactive partner.

There is an ongoing need for new click chemistry methods, particularlyfor the preparation of biologically active materials with useful anduncommon functional groups and pharmacophores. The compounds and methodsdescribed herein address these needs.

SUMMARY

A new aspect of “click” chemistry—dubbed Sulfonyl Fluoride Exchange(SuFEx)—is described herein. SuFEx is made possible by the interplaybetween the unique hydrogen-bonding requirements of the fluoride ion andthe thermodynamic and kinetic properties of fluoride bonds to sulfur(VI)and silicon centers. Click reactions rarely involve acid-base chemistry,because acid-base reactions generally exhibit low selectivity; however,SuFEx transformations are an exception. The special nature of thefluoride ion makes this possible, requiring guidance by “H⁺” or “R₃Si⁺”under strict spatial and kinetic constraints. SuFEx chemistry usesinterfacial (aqueous/organic) and homogeneous conditions to advantage.The muted polarity of the SO₂ group allows the properties of themolecules built with SO₂ linkages to be influenced to a great degree bythe motifs being connected. The resulting sulfonyl/sulfate connectortoolbox is also powerfully enhanced by another click reaction, theconjugate (Michael) addition of nucleophiles to the special electrophileethenesulfonyl fluoride (also known as ethylenesulfonyl fluoride).

The compounds described herein are analogs of biologically activematerials such as drugs, other therapeutic agents, herbicides,pesticides, antimicrobial agents, veterinary medical agents, and thelike, which include at least one —Z—X¹—(S)(O)(X²)F group, as describedbelow, generally in place of an —OH, —NH₂ or —NHR substituent of thedrug or therapeutic agent. In some cases, the —Z—X¹—(S)(O)(X²)Fsubstituent of the analog replaces another group on the drug,therapeutic or other biologically active agent, such as a —CF₃, —OCF₃,—OMe, —OEt, or halogen (e.g., Cl or Br) substituent, or a hydrogen on acarbon of the drug or therapeutic agent.

In some embodiments, a biologically active compound described herein isrepresented by Formula (I):

wherein:

Y is a biologically active organic core group comprising one or moreunsubstituted or substituted moiety selected from an aryl group, aheteroaryl aryl group, a nonaromatic hydrocarbyl group, and anonaromatic heterocyclic group, to which each Z independently iscovalently bonded;

n is 1, 2, 3, 4 or 5;

each Z independently is O, NR, or N;

when Z is O, m is 1, X¹ is a covalent bond, and the Z is covalentlybonded to an aryl or a heteroaryl moiety of Y;

when Z is NR, m is 1, X¹ is a covalent bond or CH₂CH₂, and the Z iscovalently bonded to a nonaromatic hydrocarbyl, a nonaromaticheterocyclic, an aryl, or a heteroaryl moiety of Y;

when Z is N, either (a) m is 2, X¹ is CH₂CH₂ and the Z is covalentlybonded to a nonaromatic hydrocarbyl, a nonaromatic heterocyclic, anaryl, or a heteroaryl moiety of Y; or (b) m is 1, X¹ is a covalent bondor CH₂CH₂, and the Z is a nitrogen in an aromatic or non-aromaticheterocyclic ring portion of core group Y;

each X² independently is O or NR; and

each R independently comprises H or a substituted or unsubstituted groupselected from an aryl group, a heteroaryl aryl group, a nonaromatichydrocarbyl group, and a nonaromatic heterocyclic group.

Substituents that may be present on the various hydrocarbyl, aryl,heteroaryl, and heterocyclic components of the compounds of Formula (I),include, e.g., one or more substituent selected from the groupconsisting of a hydrocarbyl moiety (e.g., an alkyl, alkenyl, alkynyl,aryl, alkylaryl, arylalkyl, or any combination of two or more thereof),—OR⁴, —N(R⁴)₂, —N⁺(R⁴)₃, —SR⁴, —OC(═O)R⁴, —N(R⁴)C(═O)R⁴, —SC(═O)R⁴,—OC(═O)OR⁵, —N(R⁴)C(═O)OR⁵, —SC(═O)OR⁵, —OC(═O)N(R⁴)₂,—N(R⁴)C(═O)N(R⁴)₂, —SC(═O)N(R⁴)₂, —OC(═O)SR⁵, —N(R⁴)C(═O)SR⁵,—SC(═O)SR⁵, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)N(R⁴)₂, —C(═O)SR⁴, —OC(═NR⁴)R⁴,—N(R⁴)C(═NR⁴)R⁴, —SO₂R⁴, —SO₂OR⁴, —SO₂(NR⁴)₂, —N(R⁴)SO₂OR⁵,—N(R⁴)SO₂N(R⁴)₂, —OSO₂OR⁵, —OSO₂N(R⁴)₂, —P(═O)(OR⁴)₂, —OP(═O)(OR⁴)₂,—OP(═O)R⁵(OR⁴), fluoro, chloro, bromo, iodo, —NO₂, —N₃, —N═N—Ar¹, —CN, aheteroaryl moiety (including heteroaryl materials comprising a singlearomatic ring, or multiple fused aromatic rings in which at least one ofthe fused rings includes a heteroatom), a nonaromatic heterocyclicmoiety, a fused 5-member nonaromatic carbocyclic ring, a fused 5-memberheterocyclic ring, a fused 6-member nonaromatic carbocyclic ring, afused 6-member nonaromatic nitrogen-containing heterocyclic ring, andany combination of two or more thereof. In the foregoing substituents,each R⁴ independently is H, hydrocarbyl, heteroaryl, or a nonaromaticheterocyclic moiety; each R⁵ independently is hydrocarbyl, heteroaryl,or a nonaromatic heterocyclic moiety; and each Ar¹ independently is arylor heteroaryl.

Certain compounds of Formula (I) can be prepared by reacting a compoundY—(ZH)_(n) with a reagent comprising a —S(O)(X²)F group, such as SO₂F₂,CH₂═CHSO₂F, and other reactions described in detail herein, including,e.g., conversion of the corresponding chloride containing molecules tothe fluoro compounds by nucleophilic displacement of Cl by F (e.g.,using a fluoride salt or a bifluoride salt). In some embodiments,Y—(ZH)_(n) is a known, commercial drug that includes one or more primaryor secondary amino substituent and/or aromatic OH substituent, which isreactive toward the reagent. In other embodiments, Y—(ZH)_(n) is ananalog of a known, commercial drug that includes one or more primary orsecondary amino substituent and/or aromatic OH substituent, which isreactive toward the reagent, in place of a hydrogen or anothersubstituent, e.g., in place of OMe, OCF₃, CF₃, halogen, etc.) of aknown, commercial drug.

In some embodiments, the compound of Formula (I) can be represented byFormula (II) or Formula (III):

wherein each X² independently is O or NR⁵. In preferred embodiments, X²is O, and the compounds can be represented by Formula (III):

In Formula (II) and Formula (III), A is a biologically active organiccore group. In some embodiments, A comprises at least one moiety, R¹,and “n” is 1, 2, 3, 4 or 5. In other words, the compounds of Formula(II) include an A group which has therapeutic/medicinal activity, inwhich one or more —ZH group (i.e., —OH, —NH₂, or —NHR) of an R^(i)moiety of A is substituted by a [—Z—(X¹—S(O)(X²)F)_(m)]_(n) group,generally by reaction with a reagent that will condense with the ZH orotherwise react with the ZH group with elimination of the H therefrom.The compounds of Formula (II) retain the therapeutic activity of thecore group A. Each Z independently is O, NR, or N; each Z is covalentlybonded to an R^(i) moiety of A; and each R independently comprises ahydrocarbyl group. When Z is O, m is 1, and each X¹ is a covalent bond.When Z is NR, m is 1, and each X¹ independently is a covalent bond orCH₂CH₂. When Z is N, m is 2, and X¹ is CH₂CH₂. Each R¹ independently isan aryl group, a heteroaryl group, or a substituted aryl group havingthe formula:

wherein x and y are 0, 1 or 2; and the sum of x and y is at least 1 whenR¹ is the substituted aryl; and each R² and R³ independently is asubstituent selected from the group consisting of a hydrocarbyl moiety,—OR⁴, —N(R⁴)₂, —N⁺(R⁴)₃, —SR⁴, —OC(═O)R⁴, —N(R⁴)C(═O)R⁴, —SC(═O)R⁴,—OC(═O)OR⁵, —N(R⁴)C(═O)OR⁵, —SC(═O)OR⁵, —OC(═O)N(R⁴)₂,—N(R⁴)C(═O)N(R⁴)₂, —SC(═O)N(R⁴)₂, —OC(═O)SR⁵, —N(R⁴)C(═O)SR⁵,—SC(═O)SR⁵, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)N(R⁴)₂, —C(═O)SR⁴, —OC(═NR⁴)R⁴,—N(R⁴)C(═NR⁴)R⁴, —SO₂R⁴, —SO₂OR⁴, —SO₂(NR⁴)₂, —N(R⁴)SO₂OR⁵,—N(R⁴)SO₂N(R⁴)₂, —OSO₂OR⁵, —OSO₂N(R⁴)₂, —P(═O)(OR⁴)₂, —OP(═O)(OR⁴)₂,—OP(═O)R⁵(OR⁴), fluoro, chloro, bromo, iodo, —NO₂, —N₃, —N═N—Ar¹, —CN, aheteroaryl moiety, a nonaromatic heterocyclic moiety, and anycombination of two or more thereof. Alternatively, an R² and an R³together form a ring selected from a fused 5-member nonaromaticcarbocyclic ring, a fused 5-member heterocyclic ring, a fused 6-membernonaromatic carbocyclic ring, and a fused 6-member nonaromaticnitrogen-containing heterocyclic ring. In the foregoing substituents,each R⁴ independently is H, hydrocarbyl, heteroaryl, or a nonaromaticheterocyclic moiety; each R⁵ independently is hydrocarbyl, heteroaryl,or a nonaromatic heterocyclic moiety; and each Ar¹ independently is arylor heteroaryl. In some embodiments, when Z is N, the Z alternatively canbe a nitrogen in an aromatic or non-aromatic heterocyclic ring portionof the core group A, in which case, m is 1, and X¹ can be a covalentbond or a —CH₂CH₂— group.

In the compounds of Formulas (II) and (III), each Ar¹, hydrocarbyl,heteroaryl, nonaromatic heterocyclic moiety, fused 5-member nonaromaticcarbocyclic ring, fused 5-member heterocyclic ring, fused 6-membernonaromatic carbocyclic ring, and fused 6-member nonaromaticnitrogen-containing heterocyclic ring independently can be unsubstitutedor can be substituted with a group comprising at least one atom otherthan carbon.

In some embodiments of the compounds of Formula (I) and Formula (II),each X² is O, each Z independently is O, NR, or N; and at least one Z isO. In some other embodiments of the compounds of Formula (I) and Formula(II), each X² is O, each Z independently is O, NR, or N; and at leastone Z is NR. In yet other embodiments of the compounds of Formula (I)and Formula (II), each X² is O, each Z independently is O, NR, or N; andat least one Z is N.

In some embodiments of the compounds of Formula (I) and Formula (II),each X² is O, each X¹ is a covalent bond, and each Z is O. In some otherembodiments of the compounds of Formula (I) and Formula (II), each X² isO, each X¹ is a covalent bond, and each Z is NR. In yet otherembodiments of the compounds of Formula (I) and Formula (II), each X² isO, each X¹ is —CH₂CH₂—, and each Z independently is NR or N.

In one embodiment of the compound represented by Formula (III), each mis 1; each Z independently is O or NR; and each X¹ is a covalent bond.

In one embodiment of the compound represented by Formula (III), each Zindependently is O, NR, or N; at least one Z is O; when Z is O, m is 1,and each X¹ is a covalent bond; when Z is NR, m is 1, and each X¹independently is a covalent bond or CH₂CH₂; and when Z is N, m is 2, andX¹ is CH₂CH₂.

In one embodiment of the compound represented by Formula (III), each Zindependently is O, NR, or N; at least one Z is NR; when Z is O, m is 1,and X¹ is a covalent bond; when Z is NR, m is 1, and each X¹independently is a covalent bond or CH₂CH₂; when Z is N, m is 2, andeach X¹ is CH₂CH₂.

In one embodiment of the compound represented by Formula (III), each Zindependently is O, NR, or N; at least one Z is N; when Z is O, m is 1,and X¹ is a covalent bond; when Z is NR, m is 1, and each X¹independently is a covalent bond or CH₂CH₂; and when Z is N, m is 2, andeach X¹ is CH₂CH₂.

In one embodiment of the compound represented by Formula (III), each mis 1; each Z is O; and each X¹ is a covalent bond.

In one embodiment of the compound represented by Formula (III), each Zis NR; and each X¹ independently is a covalent bond or CH₂CH₂.

In one embodiment of the compound represented by Formula (III), each mis 2; each Z is N; and each X¹ is CH₂CH₂.

In some embodiments of the compounds of Formulas (I), (II) and (III), Yor A comprises, e.g., an antimicrobial agent in which one or morehydrogen on an oxygen, nitrogen or a combination thereof is substitutedby [—X¹—SO₂F)_(m)]_(n); an enzyme inhibitor in which one or morehydrogen on an oxygen, nitrogen or a combination thereof is substitutedby [—X¹—SO₂F)_(m)]_(n); a medicament for treating a non-microbialdisease in which one or more hydrogen on an oxygen, nitrogen or acombination thereof is substituted by [—X¹—SO₂F)_(m)]_(n); or atherapeutic agent that targets a pathogen (e.g., an antibiotic such asvancomycin, rifamycin, rifampicin, teicoplanin, sulfacetamide,amoxicillin, novobiocin, a tetracycline compound, tetracycline,oxytetracycline, methacycline, minocycline, chlorotetracycline,doxycycline, rolitetracycline, demeclocycline, sulfanilamide,sulfamethoxazole, norfloxacin, gatifloxacin, gemifloxacin, ananti-tubercular compound, isoniazid, rifampicin, streptomycin,ciprofloxacin, moxifloxacin, aminosalicylic acid, and the like; or anprotozoal agent such as an anti-malarial agent, quinine, quinocrine,atovaquone, mefloquine, sulfadoxine, hydrochloroquine iodoquinol,paramomycin, and the like).

In some embodiments of the compounds of Formulas (I), (II) and (III), Yor A comprises, a therapeutic agent that targets an active site in ahost subject, e.g., a non-steroidal anti-inflammatory agents (NSAIDs)such as naproxen, ibuprofen, aspirin, tolmetin, flurbiprofen, sulindac,piroxicam, nabumeton, flufenamic acid, tolfenamic acid, diclofenac, andthe like; antineoplastic agents such as bleomycin, cytarabine,dacarbazine, anthracyclines (e.g., daunorubicin, doxorubicin, and thelike), epirubicin, etoposide, flutamide, gemcitabine, idarubicin,leuprolide, leuprorelin, mercaptopurine, methotrexate, mitomycin,mitoxantrone, pemetrexed, pentostatin, procarbazine, suramin,teniposide, thioguanine, thiotepa, uracil mustard (uramastine), and thelike; opiates such as morphine, buprenorphine, hydromorphone,oxymorphone, dihydromorphone, methyldihydromorphinone, butorphanol, andthe like; analgesics such as pregabalin, tetrahydrocannabinol, fentanyl,flupirtine, oxycodone, acetaminophen, salicylamide, and the like;anti-depressants such as fluoxetine (PROZAC), sertraline (ZOLOFT),duloxetine (CYMBALTA), amoxapine, maprotiline, mianserin, nomifensin,trazodine, viloxazine, aripirazole, bupropion (WELLBUTRIN),desvenlafaxine, duloxetine, paroxetine, and the like; COX 2 inhibitorssuch as celecoxib, rofecoxib, lumiracoxib, etoricoxib, firocoxib,nimesulide, and the like; COX-LOX inhibitors such as licofelone,clonidine, and the like; opioid receptor antagonists such as naltrexone,naloxone, naltrindole, and the like; Alzheimer's disease medicationssuch as epigallocatechin gallate (EGCG), memantine, galantamine, and thelike; statins such as atorvstatin (LIPITOR), rosuvastatin, and the like;erectile dysfunction medications such as sildenafil (VIAGRA), tadalafil(CIALIS), vardenafil (LEVITRA), apomorphine, and the like; anti-asthmamedications such as salbutamol (albuterol), salmeterol, terbutaline,formoterol, metaproterenol, and the like; cholinesterase inhibitors suchas edrophonium, tacrine, and the like; sympathomimetic drugs such asphenylephrine, amphetamine, methoxamine, prenalterol, terbutaline,ritodrine, and the like; anti-seizure agents such as lamotrigine,vigabatrine, gabapentin, pregabalin, and the like; neuromuscularblockers such as tubocurarine, cisatracurium, and the like; intestinalsteroid absorption inhibitors such as ezetimibe,(3R,4S)-1,4-bis(4-methoxyphenyl)-3-(3-phenylpropyl)-2-azetidinone, andthe like; endocrine drugs such as thyroxine, somatostatin, and the like;estrogenic agents, antagonists and agonists, such as raloxifene,estradiol, ethynylestradiol, diethylstilbesrol, and the like; antiviralagents such as acyclovir, valacyclovir, penciclovir, cidofovir,zalcitibine, adefovir, entacavir, and the like; anorectic agents such asphentermine, and the like; anticoagulants such as warfarin,acenocoumarol, and the like; antihypertives and beta blockers such aslisinopril, nadolol, atenolol, acebutolol, betaxolol, carvediol,esmolol, and the like; seratonin receptor agonists and seratonin uptakeinhibitors such as seratonin, sertraline, dolasetron, fluoxetine, andthe like; diuretics such as hydrochlorothiazide, bumetanide, furosemide,pinoresinol, and the like; calcium channel blockers such as amlodipinebesylate, mibefradin hydrochloride, and the like; as well as femalelibido enhancing compounds such as flibanserin(1-(2-{4-[3-(Trifluoromethyl)phenyl]piperazin-1-yl}ethyl)-1,3-dihydro-2H-benzimidazol-2-one;Sprout Pharmaceuticals). Other suitable materials include peptide-basedand amino acid-based agents, particularly tyrosine,2,6-dimethyltyrosine, lysine, and peptides comprising one or moreresidues selected from tyrosine, 2,6-dimethyltyrosine, and lysine suchas leuprolide (ENANTONE, a tyrosine-containing peptide pituitary GnRHreceptor antagonist), glatiramer (a random copolymer of lysine alanineaspartic acid and tyrosine, tradename CAPOXONE, an immunomodulator). Asis well known in the medical art, drugs within in a particularclassification (e.g., antibiotic, estrogenic agent, antineoplasticagent, and the like) may have therapeutic uses and indications for morethan one type of disease or condition.

In some embodiments, the compounds of Formulas (I), (II) and (III), Y orA are therapeutic agents that exhibit activity toward substantially thesame therapeutic target as the pharmaceutically active organic coregroup, A.

The compounds of Formulas (I), (II) and (III), Y or A that havetherapeutic activity can be formulated as a pharmaceutical compositionin combination with a pharmaceutically acceptable carrier, vehicle, ordiluent.

The present invention also provides methods for preparing the compoundsof Formula (II) and Formula (I). In one embodiment, a method ofpreparing a compound of Formula (II) or Formula (I) comprises reacting acompound of Formula (IV) or (V), respectively, with SO₂F₂ in thepresence of a base:

in which A, Y, m and n are as defined for compounds of Formulas (I) and(II), and Z is O or NR. Examples of suitable bases include (e.g., analkali metal hydroxide, such as NaOH, KOH and the like), an alkali metalalkoxide (e.g., potassium tert-butoxide, sodium methoxide, and thelike), a nitrogen base (preferably a tertiary amine, such astriethylamine or diisopropylethylamine; an amidine such as DBU(1,8-diazabicyclo[5.5.0]undec-7-ene, and the like); a guanidine such astetramethylguanidine, and the like), and the like.

In another embodiment, a method of preparing a compound of Formula (II)or Formula (I) in which each m is 2, each Z is N, and each X¹ is CH₂CH₂,comprises reacting a compound of Formula (IV) or (V), respectively, withCH₂═CH—SO₂F (“ESF”), which readily condenses with amino compoundscomprising at least one N—H bond.

The compounds of Formula (I) and Formula (II), including the compoundsof Formula (III), have a number of useful and surprisingcharacteristics. In many embodiments, the compounds have therapeuticactivity toward substantially the same therapeutic target as thecorresponding compound comprising the core group Y or A, but with one ormore useful additional properties of characteristics, such as enhancedsolubility, enhanced bioavailability, ability to covalently link with atarget group in a subject or pathogen, a lower water/octanol partitioncoefficient (Log P) than the parent compound (particularly when —OSO₂Freplaces a —OCF₃ group in the parent therapeutic compound), andproviding a handle for selectively attaching the compound of Formula(II) to a useful group such as a dye (e.g., a fluorescent dye), biotin,a polymer (e.g., a polystyrene resin or other conventional polymer, aswell as polymers described in commonly owned copending PCT ApplicationSerial No. PCT/US2013/072871, which is incorporated herein by referencein its entirety) by displacement of F. The —OSO₂F group, can beconsidered as a bioisostere for —OCF₃, making the —OSO₂F group anattractive replacement for —OCF₃, particularly if a lower Log P isdesired.

In addition to biological activity, per se, Ar—OSO₂F groups provide aneffective protecting group or precursor for ArOH and ArOSO₃ ⁻ compounds.The SO₂F moiety, in the various forms described herein also provides ahandle for covalent attachment of organic compounds to substrates thatbear a phenolic OH group, an amino group, and the like, e.g., attachmentto a surface, such as a modified polystyrene bead or a glass surface, orattachment to another molecule, such as a protein (e.g., via anucleophilic side chain of an amino acid residue in the active site of aprotein). The —OSO₂F group also can be useful as a leaving group innucleophilic aromatic substitution reactions, or other displacement orcoupling reactions in place of, e.g., a halogen group, a triflate, amesylate, or other leaving group. In addition, the biologically activeSO₂F compounds described herein can be utilized as screening agents toidentify drug targets, e.g., by covalent binding to the active site forthe drugs.

The following non-limiting listing illustrates various embodiments ofthe compounds, compositions, methods and uses described herein:

Embodiment 1 is a compound represented by Formula (I):

wherein:

Y is a biologically active organic core group comprising one or moreunsubstituted or substituted moiety selected from an aryl group, aheteroaryl aryl group, a nonaromatic hydrocarbyl group, and anonaromatic heterocyclic group, to which each Z independently iscovalently bonded;

n is 1, 2, 3, 4 or 5;

each Z independently is O, NR, or N;

when Z is O, m is 1, X¹ is a covalent bond, and the Z is covalentlybonded to an aryl or heteroaryl moiety of Y;

when Z is NR, m is 1, X¹ is a covalent bond or CH₂CH₂, and the Z iscovalently bonded to a nonaromatic hydrocarbyl, a nonaromaticheterocyclic, an aryl, or heteroaryl moiety of Y;

when Z is N, either (a) m is 2, X¹ is CH₂CH₂ and the Z is covalentlybonded to a nonaromatic hydrocarbyl, a nonaromatic heterocyclic, anaryl, or a heteroaryl moiety of Y; or (b) m is 1, X¹ is a covalent bondor CH₂CH₂, and the Z is a nitrogen in an aromatic or non-aromaticheterocyclic ring portion of core group Y;

each X² independently is O or NR; and

each R independently comprises H or a substituted or unsubstituted groupselected from an aryl group, a heteroaryl aryl group, a nonaromatichydrocarbyl group, and a nonaromatic heterocyclic group.

Embodiment 2 is a compound of Embodiment 1, wherein each X² is O, each Zindependently is O, NR, or N; and at least one Z is O.

Embodiment 3 is a compound of Embodiment 1, wherein, each X² is O, eachZ independently is O, NR, or N; and at least one Z is NR.

Embodiment 4 is a compound of Embodiment 1, wherein, each X² is O, eachZ independently is O, NR, or N; and at least one Z is N.

Embodiment 6 is a compound of Embodiment 1, wherein, each X² is O, eachX¹ is a covalent bond, and each Z is NR.

Embodiment 7 is a compound of Embodiment 1, wherein, each X² is O, eachX¹ is —CH₂CH₂—, and each Z independently is NR or N.

Embodiment 8 is a compound of any one of Embodiments 1 to 7, wherein atleast one Z is covalently bonded to a heteroaryl moiety of Y.

Embodiment 9 is a compound of any one of Embodiments 1 to 8, wherein atleast one Z is covalently bonded to an aryl moiety of Y.

Embodiment 10 is a compound of any one of Embodiments 1 to 9, wherein atleast one Z is covalently bonded to a non-aromatic carbon of Y.

Embodiment 11 is a compound of any one of Embodiments 1 to 10, whereinone or more of the aryl, heteroaryl aryl, nonaromatic hydrocarbyl, ornonaromatic heterocyclic portions of the compounds of Formula (I),include one or more substituent selected from a hydrocarbyl moiety,—OR⁴, —N(R⁴)₂, —N⁺(R⁴)₃, —SR⁴, —OC(═O)R⁴, —N(R⁴)C(═O)R⁴, —SC(═O)R⁴,—OC(═O)OR⁵, —N(R⁴)C(═O)OR⁵, —SC(═O)OR⁵, —OC(═O)N(R⁴)₂,—N(R⁴)C(═O)N(R⁴)₂, —SC(═O)N(R⁴)₂, —OC(═O)SR⁵, —N(R⁴)C(═O)SR⁵,—SC(═O)SR⁵, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)N(R⁴)₂, —C(═O)SR⁴, —OC(═NR⁴)R⁴,—N(R⁴)C(═NR⁴)R⁴, —SO₂R⁴, —SO₂OR⁴, —SO₂(NR⁴)₂, —N(R⁴)SO₂OR⁵,—N(R⁴)SO₂N(R⁴)₂, —OSO₂OR⁵, —OSO₂N(R⁴)₂, —P(═O)(OR⁴)₂, —OP(═O)(OR⁴)₂,—OP(═O)R⁵(OR⁴), fluoro, chloro, bromo, iodo, —NO₂, —N₃, —N═N—Ar¹, —CN, aheteroaryl moiety (including heteroaryl materials comprising a singlearomatic ring, or multiple fused aromatic rings in which at least one ofthe fused rings include a heteroatom), a nonaromatic heterocyclicmoiety, a fused 5-member nonaromatic carbocyclic ring, a fused 5-memberheterocyclic ring, a fused 6-member nonaromatic carbocyclic ring, afused 6-member nonaromatic nitrogen-containing heterocyclic ring, andany combination of two or more thereof, each R⁴ independently is H,hydrocarbyl, heteroaryl, or a nonaromatic heterocyclic moiety; each R⁵independently is hydrocarbyl, heteroaryl, or a nonaromatic heterocyclicmoiety; and each Ar¹ independently is aryl or heteroaryl.

Embodiment 12 is a compound of any one of Embodiments 1 to 11, whereinthe compound is an estrogenic steroid that includes at least one—Z—X¹—(S)(O)(X²)F group. As used herein, “estrogenic steroid” refers tosteroids in which the “A-ring” of the archetypical tetracyclic steroidstructure is aromatic, e.g., as illustrated below:

in which R generally is H or alkyl, R′ typically is OH, and R″ typicallyis H or ethynyl, and which can be substituted on any of the A, B, C, orD rings thereof or on R by one or more substituent such as describedherein with respect to embodiment 11, above, and the —Z—X¹—(S)(O)(X²)Fgroup can be present on any of the A, B, C, or D rings, on R, R′ R″ oranother substituent thereof, in place of OR, R′ or R″. Non-limitingexamples of such estrogenic steroids include, e.g., estradiol, estrone,estriol, ethynylestradiol, and the like.

Embodiment 13 is a compound of any one of Embodiments 1 to 11, whereinthe compound is a corticosteroid that includes at least one—Z—X¹—(S)(O)(X²)F group. As used herein, “corticosteroid” refers tosteroids having the archetypical corticosteroid structure illustratedbelow:

in which R typically is H, R′ generally is OH or oxo (═O), and R″typically is —CH₂OH, and which can be substituted on any of the A, B, C,or D rings thereof or on R, R′ or R″, by one or more substituent such asdescribed herein with respect to embodiment 11, above, and the—Z—X¹—(S)(O)(X²)F group can be present on any of the A, B, C, or Drings, on another substituent attached to one of the rings, on an R, R′or R″ group, or in place of an R, R′ or R″ group, (e.g., attached to anaryl or heteroaryl substituent when Z is O). Non-limiting examples ofsuch corticosteroids include, e.g., cortisone, hydrocortisone,prednisone, prednisolone, triamcinolone, methyl prednisolone,prednylidene, fluocortilone, parametasone, dexamethasone, betamethasone,and the like.

Embodiment 14 is a compound of any one of Embodiments 1 to 11, whereinthe compound is an amphetamine compound that includes at least one—Z—X¹—(S)(O)(X²)F group. As used herein, “amphetamine compound” refersto compounds which include the archetypal amphetamine core substructure:

and which can be substituted on any portion thereof by one or moresubstituent such described herein with respect to Embodiment 11, above,and the —Z—X¹—(S)(O)(X²)F group can be present on any portion of theamphetamine core or can be covalently attached to another substituent onthe amphetamine core. Non-limiting examples of such amphetaminecompounds include, e.g., psychostimulants such as amphetamine,metamphetamine, amfetaminil, fenetylline, methylphenidate, prolintane;and anorectics such as cathine (norpseudoephedrine), amfepramone,mefanorex, fenfluramine, and the like.

Embodiment 15 is a compound of any one of Embodiments 1 to 11, whereinthe compound is benzodiazepine compound that includes at least one—Z—X¹—(S)(O)(X²)F group. As used herein, “benzodiazepine compound”refers to compounds which include the archetypal benzodiazepine coresubstructure:

in which R′ typically is a halogen or nitro group, R″ typically is anoxo (═O) or NR group, and which can be substituted on any portionthereof by one or more substituent such as described herein with respectto Embodiment 11, above, and the —Z—X¹—(S)(O)(X²)F group can be presenton any portion of the benzodiazepine core or can be covalently attachedto another substituent on the benzodiazepine core, or in place of R′ orR″. Non-limiting examples of such benzodiazepine compounds include,e.g., chlorodiazepine, demoxepam, chlordiazepoxide, diazapam, prazepam,oxazepam, dipotassoim chlorazepate, lorazepam, clonazepam, bromazepam,clobazam, temazepam, flurazepam, lormetazepam, nitrazepam, and the like.

Embodiment 16 is a compound of any one of Embodiments 1 to 11, whereinthe compound is a barbiturate compound that includes at least one—Z—X¹—(S)(O)(X²)F group. As used herein, “barbiturate compound” refersto compounds which include the archetypal barbiturate core substructure:

in which the R groups can be substituent such as described in Embodiment11, above, and the —Z—X¹—(S)(O)(X²)F group can be present on any portionof the benzodiazepine core or can be covalently attached to anothersubstituent on the amphetamine core, or in place of an R group.Non-limiting examples of such barbiturate compounds include, e.g., suchas vinylbital, aprobarbital, secbutabarbital, pentobarbital,cyclobarbital, phenobarbital, and the like.

Embodiment 17 is a compound of any one of Embodiments 1 to 11, whereinthe compound is a morphine derivative that includes at least one—Z—X¹—(S)(O)(X²)F group. As used herein, “morphine derivative” refers tocompounds which include the archetypal morphine core substructure:

in which R typically is hydroxyl or alkoxy, R′ typically is OH or oxo,and the compound can be substituted by a substituent such described inEmbodiment 11 on any portion of the core structure or on included on orin place of R or R′, and, and the —Z—X¹—(S)(O)(X²)F group can be presenton any portion of the morphine core or can be covalently attached toanother substituent on the morphine core or in place of R or R′.Non-limiting examples of such morphine derivatives include, e.g.,morphine, codeine, diamorphine, dihydrocodeine, hydromorphone,hydrocodone, oxycodone, oxymorphone, levorphanol, and the like.

Embodiment 18 is a compound of any one of Embodiments 1 to 11, whereinthe compound is penam antibiotic that includes at least one—Z—X¹—(S)(O)(X²)F group. As used herein, “penam antibiotic” refers to anantibiotic comprising the archetypical penam core structure:

and which can be substituted on any portion thereof by one or moresubstituent such described herein with respect to Embodiment 11, above,and the —Z—X¹—(S)(O)(X²)F group can be present on any portion of thepenam core or can be covalently attached to another substituent on thepenam core. Non-limiting examples of such penam antibiotics include,e.g., penicillin, benzylpenicillin (penicillin G),phenoxymethylpenicillin (penicillin V), oxacillin, dicloxacilin,flucloxacillin, ampicillin, amoxicillin, epicillin, azlocillin,mezlocillin, piperacillin, apalcillin, carbenicillin, ticarcillin,temocillin, and the like.

Embodiment 19 is a compound of any one of Embodiments 1 to 11, whereinthe compound is a cephem antibiotic that includes at least one—Z—X¹—(S)(O)(X²)F group. As used herein, “cephem antibiotic” refers toan antibiotic comprising the archetypical cephem core structure:

and which can be substituted on any portion thereof by one or moresubstituent such described herein with respect to Embodiment 11, above,and the —Z—X¹—(S)(O)(X²)F F group can be present on any portion of thecephem core or can be covalently attached to another substituent on thecephem core. Non-limiting examples of such cephem antibiotics include,e.g., cephalosporin, cefalotin, cefazolin, cefazedone, cefamandole,cefuroxime, cefotiam, cefotaxime, ceftizoxime, ceftriaxone, ceftazidime,cefoperazone, cefixime, cefmetazole, cefonicid, cefapirin, ceforanide,cefalexin, cefaclor, cefradine, cefadroxil, and the like.

Embodiment 20 is a compound of any one of Embodiments 1 to 11, whereinthe compound is a carbapenem antibiotic that includes at least one—Z—X¹—(S)(O)(X²)F group. As used herein, “carbapenem antibiotic” refersto an antibiotic comprising the archetypical carbapenem core structure:

and which can be substituted on any portion thereof by one or moresubstituent such described herein with respect to embodiment 11, above,and the —Z—X¹—(S)(O)(X²)F group can be present on any portion of thecarbopenem core or can be covalently attached to another substituent onthe carbapenem core. Non-limiting examples of such carbapenemantibiotics include, e.g., thienamycin, imipenem, meropenem, ertapenem,doripenem, biapenem, razupenem, tebipenem, lenapenem, and tomopenem.

Embodiment 21 is a compound of any one of Embodiments 1 to 11, whereinthe compound is a tetracycline antibiotic that includes at least one—Z—X¹—(S)(O)(X²)F group. As used herein, “tetracycline antibiotic”refers to an antibiotic comprising the archetypical tetracycline corestructure:

in which R typically is a hydrogen, halogen (e.g., Cl), dialkylamino(e.g., dimethylamino), R′ and R″ typically are hydrogen, hydroxyl ormethyl, R′″ typically is hydrogen or hydroxyl, and which can besubstituted on any portion thereof by one or more substituent such asdescribed herein with respect to Embodiment 11, above, and the—Z—X¹—(S)(O)(X²)F group can be present on any portion of thetetracycline core or can be covalently attached to another substituenton the tetracycline core, or in place of R, R′, R″ or R′″. Non-limitingexamples of such tetracycline antibiotic compounds include, e.g.,tetracycline, oxytetracycline, demiclocycline, doxycycline, minocycline,and rolitetracycline.

Embodiment 22 is a compound of any one of Embodiments 1 to 11, whereinthe compound is a quinolone antibiotic that includes at least one—Z—X¹—(S)(O)(X²)F group. As used herein, “quinolone antibiotic” refersto an antibiotic comprising the archetypical quinolone core structure:

in which X typically is C or N, R typically is a hydrogen or methyl, R′typically is hydrogen or F, and R″ typically is hydrogen when X is C andabsent when X is N, R′″ typically is alkyl (e.g., ethyl) or cycloalkyl(e.g., cyclopropyl), and which can be substituted on any portion thereofby one or more substituent such as described herein with respect toEmbodiment 11, above, and the —Z—X¹—(S)(O)(X²)F group can be present onany portion of the quinolone core or can be covalently attached toanother substituent on the quinolone core, or in place of R, R′, R″ orR′″. Non-limiting examples of such quinolone antibiotic compoundsinclude, e.g., norfloxacine, ciprofloxacin, and enoxacine.

Embodiment 23 is a compound of any one of Embodiments 1 to 11, whereinthe compound is macrolide antibiotic that includes at least one—Z—X¹—(S)(O)(X²)F group. As used herein, “macrolide antibiotic” refersto an antibiotic having a macrocyclic lactone core structure. In someembodiments the macrolide antibiotic has an archetypal erythromycin-typecore structure:

in which R typically is a glycoside (sugar) group, R′ is anaminoglycoside (amino sugar) group, and R′″ typically is ethyl. Themacrolide antibiotics can be substituted on any portion thereof by oneor more substituent such as described herein with respect to Embodiment11, above, and the —Z—X¹—(S)(O)(X²)F group can be present on any portionof the macrolide core or can be covalently attached to anothersubstituent on the macrolide core. Non-limiting examples of suchmacrolide antibiotic compounds include, e.g., azithromycin,clarithromycin, erythromycin, fidaxomicin, telithromycin, carbomycin a,josamycin, kitasamycin, oleandomycin, solithromycin, spiramycin,roleandomycin, and the like.

Embodiment 24 is a compound of any one of Embodiments 1 to 11, whereinthe compound is an aminoglycoside antibiotic that includes at least one—Z—X¹—(S)(O)(X²)F group. As used herein, “aminoglycoside antibiotic”refers to an oligosaccharide (typically a trisaccharide ortetrasaccharide) antibiotic comprising at least one amino sugarcomponent (e.g., streptamine or 2-desoxystreptamine) in theoligosaccharide chain thereof. The aminoglycoside antibiotics can besubstituted on any portion thereof by one or more substituent such asdescribed herein with respect to Embodiment 11, above, and the—Z—X¹—(S)(O)(X²)F group can be present on any portion of theaminoglycoside core or can be covalently attached to another substituenton the aminoglycoside core. Non-limiting examples of such aminoglycosideantibiotics, include, e.g., streptomycin, neomycin B, gentamicin,kanamycin, and the like.

Embodiment 25 is a compound of any one of Embodiments 1 to 11, whereinthe compound is a transthyretin (TTR) binding compound represented byFormula (VI) or Formula (VIa):

in which each R is alkyl (e.g., methyl, ethyl, propyl) or halogen (e.g.,Cl, Br), L′ is as shown (i.e., trans vinyl, diazo, or1O,3N,4N-oxadiazol-2,5-diyl, and the —OSO₂F group in each formula can bebonded to the 3, 4, or 5 position on the “A” ring of the compound. Someof the compounds of Formula (VI) and (VIa) can irreversibly bind to theTTR binding site, stabilizing the TTR protein tertiary structure.

Embodiment 26 is a compound of any one of Embodiments 1 to 11, whereinthe compound is an analog of a biologically active material selectedfrom an antimicrobial agent, an enzyme inhibitor, a medicinal agenthaving activity for treating a non-microbial disease, a medicinal agenttargets a pathogen, an antibiotic, an anti-protozoal agent, and atherapeutic agent that targets an active site in a host subject; whichanalog includes at least one —Z—X¹—(S)(X²)F group.

Embodiment 27 is a compound of Embodiment 26, wherein the antibiotic isselected from vancomycin, rifamycin, rifampicin, teicoplanin,sulfacetamide, amoxicillin, novobiocin, a tetracycline compound,tetracycline, oxytetracycline, methacycline, minocycline,chlorotetracycline, doxycycline, rolitetracycline, demeclocycline,sulfanilamide, sulfamethoxazole, norfloxacin, gatifloxacin,gemifloxacin, trimethoprim, pyrimethamine, cefadroxil, ananti-tubercular compound, isoniazid, rifampicin; streptomycin,ciprofloxacin, moxifloxacin, and aminosalicylic acid.

Embodiment 28 is a compound of Embodiment 26, wherein the anti-protozoalagent is an anti-malarial agent selected from quinine, quinocrine,atovaquone, mefloquine, sulfadoxine, hydrochloroquine iodoquinol, andparamomycin.

Embodiment 29 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises anon-steroidal anti-inflammatory agent (NSAID) selected from naproxen,ibuprofen, aspirin, tolmetin, flurbiprofen, sulindac, piroxicam,nabumeton, flufenamic acid, tolfenamic acid, and diclofenac.

Embodiment 30 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises anantineoplastic agent selected from bleomycin, cytarabine, dacarbazine,an anthracycline, daunorubicin, doxorubicin, epirubicin, etoposide,flutamide, gemcitabine, idarubicin, leuprolide, mercaptopurine,methotrexate, mitomycin, mitoxantrone, pemetrexed, pentostatin,procarbazine, suramin, teniposide, thioguanine, thiotepa, and uracilmustard (uramastine).

Embodiment 31 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises an opiateselected from buprenorphine, hydromorphone, oxymorphone,dihydromorphone, and methyldihydromorphinone.

Embodiment 32 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises ananalgesic selected from pregabalin, tetrahydrocannabinol, fentanyl,flupirtine, oxycodone, acetaminophen, and salicylamide.

Embodiment 33 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises ananti-depressant selected from fluoxetine, sertraline, duloxetine,amoxapine, maprotiline, mianserin, nomifensin, trazodine, viloxazine,aripirazole, bupropion, desvenlafaxine, duloxetine, and paroxetine.

Embodiment 34 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises a COX 2inhibitor selected from celecoxib, rofecoxib, lumiracoxib, etoricoxib,firocoxib, and nimesulide.

Embodiment 35 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises a COX-LOXinhibitor selected from licofelone, and clonidine.

Embodiment 36 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises a opioidreceptor antagonist selected from naltrexone, naloxone, and naltrindole.

Embodiment 37 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises anAlzheimer's disease medication selected from epigallocatechin gallate(EGCG), memantine, and galantamine.

Embodiment 38 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises a statinselected from atorvastatin and rosuvastatin.

Embodiment 39 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises anerectile dysfunction medication selected from sildenafil, tadalafil,vardenafil, and apomorphine.

Embodiment 40 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises ananti-asthma medication selected from salbutamol, salmeterol,terbutaline, formoterol, and metaproterenol.

Embodiment 41 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises acholinesterase inhibitor selected from edrophonium and tacrine.

Embodiment 42 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises asympathomimetic drug selected from phenylephrine, amphetamine,methoxamine, prenalterol, terbutaline, and ritodrine.

Embodiment 43 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises ananti-seizure agent selected from lamotrigine and vigabatrine.

Embodiment 44 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises aneuromuscular blocker selected from tubocurarine and cisatracurium.

Embodiment 45 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises anintestinal steroid absorption inhibitor selected from ezetimibe and(3R,4S)-1,4-bis(4-methoxyphenyl)-3-(3-phenylpropyl)-2-azetidinone.

Embodiment 46 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises anendocrine drug selected from thyroxine and somatostatin.

Embodiment 47 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises anestrogenic agent, agonist or antagonist selected from raloxifene,estradiol, ethynylestradiol, and diethylstilbestrol.

Embodiment 48 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises ananti-viral agent selected from acyclovir, valacyclovir, penciclovir,cidofovir, zalcitibine, adefovir, and entacavir.

Embodiment 49 is a compound of Embodiment 26, wherein the therapeuticagent that targets an active site in a host subject comprises aseratonin receptor agonist selected from dolasetron and seratonin.

Embodiment 50 is a compound of any one of Embodiments 1 to 11, whereinthe compound is an analog of tyrosine, 2,6-dimethyltyrosine, or apeptide comprising one or more residues selected from tyrosine and2,6-dimethyltyrosine, in which the phenolic OH of the tyrosine or2,6-dimethyltyrosine is substituted by —OSO₂F.

Embodiment 51 is a compound of Embodiment 50, wherein the compound isO-fluorosulfonyltyrosine or O-fluorosulfonyl-2,6-dimethyltyrosine.

Embodiment 52 is a compound of Embodiment 50, wherein the peptide isselected from leuprolide and glatiramer, and is modified to include a—OSO₂F group in place of the phenolic OH of a tyrosine residue thereof.

Embodiment 53 is a compound of any one of Embodiments 1 to 11, whereinthe compound comprises an analog of amino acid comprising a nucleophilicside chain or a peptide comprising one or more amino acid residuecomprising a nucleophilic side chain, which includes an SO₂F or—CH₂CH₂SO₂F group in place of a hydrogen on a hydroxyl or aminosubstituent of the nucleophilic side chain.

Embodiment 54 is a compound of Embodiment 53, wherein the amino acid isselected from lysine, serine, tyrosine, histidine, and arginine.

Embodiment 55 is a compound of Embodiment 53, wherein the compound is apeptide comprising the amino acid residue selected from the groupconsisting of lysine, serine, tyrosine, histidine, and arginine, whichincludes an SO₂F or —CH₂CH₂SO₂F group in place of a hydrogen on ahydroxyl or amino substituent of the nucleophilic side chain.

Embodiment 56 is a compound of any one of Embodiments 1 to 55, whereinthe compound of Formula (I) has biological activity toward substantiallythe same target as the biologically active core group Y.

Embodiment 57 is a compound of any one of Embodiments 1 to 56, whereinthe fluorine (F) of one or more of the —Z—X¹—(S)(O)(X²)F groups thereofis enriched in ¹⁸F.

Embodiment 58 is a pharmaceutical composition comprising a compound ofany one of Embodiments 1 to 57, and a pharmaceutically acceptablecarrier, vehicle, or diluent.

Embodiment 59 is a method of preparing a compound of Embodiment 1, inwhich at least one Z thereof is O; the method comprising reacting aprecursor bearing an aromatic and/or heteroaromatic OH substituent withSO₂F₂ in the presence of a base to replace the hydrogen of the aromaticand/or heteroaromatic OH with SO₂F.

Embodiment 60 is a method of preparing a compound of Embodiments 1, inwhich at least one Z thereof is NR; the method comprising reacting aprecursor bearing an NHR substituent with SO₂F₂ in the presence of abase to replace the hydrogen of the NHR with SO₂F.

Embodiment 61 is a method of preparing a compound of Embodiments 1, inwhich at least one Z thereof is N or NR; the method comprising reactinga precursor bearing an NH₂ or NHR substituent with CH₂═CH—SO₂F by aMichael addition to replace the hydrogens of the NH₂ or the hydrogen ofthe NHR with —CH₂CH₂—SO₂F.

Embodiment 62 is a compound of any one of Embodiments 1 to 57 fortreating a disease or condition.

Embodiment 63 is the use of a compound of any one of Embodiments 1 to57, for treating a disease or condition.

Embodiment 64 is the use of a compound of any one of Embodiments 1 to57, for the preparation of a medicament for treating a disease orcondition.

Embodiment 65 is the use of a library comprising a plurality of thecompounds of any one of Embodiments 1 to 57 in a screening assay againsta biologically active receptor protein.

Embodiment 66 is a method for preparing a compound of Embodiment 57,comprising treating a compound of any one of Embodiments 1 to 56 withbifluoride ion enriched in ¹⁸F to replace at least a portion of F in thecompound with ¹⁸F.

Embodiment 67 is an-amino protected O-fluorosulfonyl-L-tyrosine offormula:

wherein “Fmoc” represents a 9-fluorenylmethyloxycarbonyl protectinggroup.

Embodiment 68 is the use of a compound of Embodiment 67 for thepreparation of a peptide or protein comprising anO-fluorosulfonyl-L-tyrosine residue.

Embodiment 69 is a peptide or protein comprising anO-fluorosulfonyl-L-tyrosine residue.

Embodiment 70 is a polypeptide of Embodiment 69, wherein the polypeptideis selected from an analog of oxytocin, indolicin, thymopentin, andarginine vassopressin, in which the tyrosine residue thereof is replacedby an O-fluorosulfonyl-L-tyrosine residue.

Embodiment 71 is the use of a compound of Embodiment 57 as an imagingagent for positron emission tomography.

Embodiment 72 is the use of a compound of any one of Embodiments 1 to 57or a peptide or protein of Embodiment 69 for covalently binding of thecompound to an active site in a receptor molecule.

Embodiment 73 is a method of preparing a sulfated polypeptide comprisingcontacting the peptide of Embodiment 69 with cesium carbonate and asolution of ammonia in methanol to selectively hydrolyze the fluorogroup of a fluorosulfonyl-L-tyrosine residue thereof and form a sulfatedtyrosine residue therefrom.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows reactions illustrating the properties of sulfonyl fluoridesvs. other sulfonyl halides. (A) resistance of ArSO₂F toward bothoxidation and reduction; (B) greater stability of sulfonyl fluoridetoward thermolysis; (C, D) chlorination vs. desired sulfonylation inreactions with ester enolates and under Friedel-Crafts conditions; (E,F) greater reactivity of acyl chloride and benzylic bromide compared tosulfonyl fluoride under non-activating conditions; (G) the power ofwater in activating sulfonyl fluoride reactivity.

FIG. 2 illustrates the essential role of fluoride stabilization andbifluoride attack in SuFEx chemistry.

FIG. 3 illustrates common methods for the synthesis of aryl (top) andalkyl (bottom) sulfonyl chlorides and fluorides. The C—S bonds of thesederivatives can be formed by nucleophilic attack of S(IV) on organicelectrophiles or attack of organic nucleophiles on electrophilic S(VI)centers.

FIG. 4 illustrates the special reactivity of bifluoride ion at awater-organic interface. [FHF]⁻ molecules at the surface lose the keyH-bonding interactions with water that stabilize this species in thebulk. As a result, bifluoride at the surface or interface is far morenucleophilic. Interactions with ArSO₂Cl leading to substitution areshown. M=the counterion for bifluoride, usually K⁺. Also shown is the[H₂F₃]⁻ ion, which is present in significant quantities along withbifluoride.

FIG. 5 provides examples of sulfonyl fluorides made with potassiumbifluoride.

FIG. 6 illustrates alkyl (top) and aryl (bottom) sulfonyl fluorides madefrom sulfonic acids. (a) NaN₃, acetone, H₂O, reflux, 8 hours; (b)(COCl)₂, CH₂Cl₂, DMF (cat.), room temperature (RT), 18 hours; (c) KFHF(sat.), CH₃CN, RT, 6 hours. (d) Na₂SO₃ (1 equiv.), H₂O, 95° C., 16 h.

FIG. 7 shows sulfonimidoyl and sulfamoyl fluorides prepared from thecorresponding chlorides. The acidic workup in reaction C is required tohydrolyze the silver acetylide formed under these conditions.

FIG. 8 illustrates small connector molecules allowing the installationof sulfonyl fluorides onto other functional structures.

FIG. 9 illustrates synthesis (top) and use (bottom) of ESF in thedecoration of nitrogen, oxygen, and carbon nucleophiles. Reactionconditions: (A) ESF, 95:5 EtOH:H₂O, 5 minutes to hours; (B) ESF, solvent(usually CH₂Cl₂ or THF), 5 min to hours; (C) ESF, PR₃ (10 mol %),CH₂Cl₂, 24 hours; (D) ESF, AcOH, reflux, 2 hours; (E) ESF, Bu₄NF (10 mol%), THF; (F) ESF, quinine (10 mol %), CH₂Cl₂.

FIG. 10 illustrates dual modes of reactivity of fluorosulfates.

FIG. 11 shows aryl fluorosulfates prepared by a convenient procedurewith gaseous SO₂F₂, in the presence of the following bases: (A) Et₃N inCH₂Cl₂, (B) Et₃N or iPr₂NEt in biphasic mixture (CH₂Cl₂/water), (C) NaHin THF, (D) DBU in MeCN.

FIG. 12 illustrates DBU-mediated conversions of aryl silyl ethers tofluorosulfates and diarylsulfates. The dotted lines in reaction B aremeant to show connectivity, not mechanism.

FIG. 13 illustrates aryl fluorosulfates in Pd-catalyzed couplingreactions.

FIG. 14 illustrates preparations of enol fluorosulfates.

FIG. 15 illustrates, in Panels B-D, preparations of N-monosubstitutedsulfamoyl fluorides; Panel A provides a comparison with the directreaction of primary amines with SO₂F₂, which does not result inN-monosubstituted sulfamoyl fluoride formation.

FIG. 16 illustrates formation of N-disubstituted sulfamoyl fluorides,with selected examples. Yields are of analytically pure materialisolated after extraction. (a) DMAP (30 mol %), MgO (5 equiv.), 4/1CH₂Cl₂/H₂O, RT, 18 hours.

FIG. 17 provides (Top) example of sulfamoyl fluoride substitution bysecondary amine; (Bottom) examples of transformations performed in thepresence of the sulfamoyl fluoride moiety.

FIG. 18 graphically illustrates loss of sulfonimidoyl fluorides as afunction of pH and nitrogen substituent.

FIG. 19 provides examples of sulfonyl fluorides made from thecorresponding chlorides using potassium bifluoride.

FIG. 20 illustrates structures of antibiotic compounds andfluorosulfonated derivatives thereof which were evaluated for activityagainst E. coli and B. subtilis.

FIG. 21 graphically illustrates reactions of fluorosulfates and sulfonylfluorides with nucleophilic amino acid side chains in receptor activesites.

FIG. 22 schematically illustrates an screening assay embodiment.

FIG. 23 schematically illustrates reactions of fluorosulfates withmultiple nucleophilic amino acid side chains in a receptor active site.

FIG. 24 schematically illustrates reactions of fluorosulfates withmultiple thiol amino acid side chains in a receptor active site.

FIG. 25 illustrates work flow of SILAC identification of labeled proteintargets using aryl-SF and aryl-OSF probes. High heavy/light ratio forFABP5 and CRABP2 indicates that they are covalently labeled by SF andOSF probes.

FIG. 26 shows recombinant FABP5 and CRABP2 were incubated with SF-3 andOSF-4 and the site of modification was identified by tandem massspectrometry. The tyrosine residues in the Arg-Tyr-Arg modules aremodified. Mutations of the tyrosine and arginine residues prevent orsignificantly impair the modification event.

FIG. 27 illustrates competition experiments using SF and OSF probessuggest chemoselective labeling. Covalent inhibitor SF-3-Cl (A) andOSF-4-E (B) could out-compete the labeling of FABP5/CRABP2 in live HeLacells by SF-3 probe (A) or OSF-4 (B), respectively. Non-covalentinhibitors all-trans retinoic acid (RA), aP2 inhibitor BMS 309403 (BMS)or SOAT2 inhibitor Avamisibe could out compete the selective labeling ofFABP5/CRABP2 in live HeLa cells by aryl-SF and aryl-OSF probes (C).

FIG. 28 illustrates additional examples of biologically active moleculesthat can be reacted with ESF.

FIG. 29 illustrates additional examples of biologically active moleculesthat can be reacted with ESF.

FIG. 30 illustrates additional examples of biologically active moleculesthat can be reacted with ESF.

FIG. 31 illustrates additional examples of biologically active moleculesthat can be reacted with ESF.

FIG. 32 illustrates additional examples of biologically active moleculesthat can be reacted with ESF.

FIG. 33 illustrates additional examples of biologically active moleculesthat can be reacted with ESF.

FIG. 34 illustrates additional examples of biologically active moleculesthat can be reacted with ESF.

FIG. 35 illustrates additional examples of biologically active moleculesthat can be reacted with ESF.

FIG. 36 illustrates additional examples of biologically active moleculesthat can be reacted with SO₂F₂.

FIG. 37 illustrates additional examples of biologically active moleculesthat can be reacted with SO₂F₂.

DETAILED DESCRIPTION

The term “alkyl” as used herein denotes saturated hydrocarbon moieties.Preferably, an alkyl group comprises 1 to 20 carbon atoms in theprincipal chain (e.g., 1 to 12 carbon atoms) and e.g., up to 30 totalcarbon atoms. These moieties may be straight or branched chain andinclude methyl, ethyl, propyl, isopropyl, butyl, hexyl, octyl, and thelike groups.

The term “alkenyl” as used herein denotes a univalent hydrocarbon groupcontaining a double bond. Preferably, alkenyl groups comprise 2 to 20carbon atoms (e.g., 2 to 12 carbon atoms) in the principal chain, and upto 30 total carbon atoms. The alkenyl groups may be straight or branchedchain, or cyclic, and include ethenyl, propenyl, isopropenyl, butenyl,isobutenyl, hexenyl, octenyl, oleyl, and the like.

The term “alkynyl” as used herein denotes a univalent hydrocarbon groupcontaining a triple bond. Preferably, alkynyl groups comprise 2 to 20carbon atoms (e.g., 2 to 12 carbon atoms) in the principal chain, and upto 30 total carbon atoms. The alkynyl groups may be straight or branchedchain, and include ethynyl, propynyl, butynyl, isobutynyl, hexynyl,octynyl, and the like.

The term “aromatic” as used herein denotes chemical compounds or groupsthat contain conjugated planar ring systems with delocalized pi electronclouds instead of discrete alternating single and double bonds. The term“aromatic” encompasses the “aryl” and “heteroaryl” groups defined below.

The terms “aryl” or “Ar” as used herein alone or as part of anothergroup denote optionally substituted homocyclic aromatic groups,preferably monocyclic or bicyclic groups containing from 6 to 12 carbonsin the ring portion, such as phenyl, biphenyl, naphthyl, anthracenyl,substituted phenyl, substituted biphenyl or substituted naphthyl.

The term “heteroaryl” as used herein alone or as part of another groupdenote optionally substituted aromatic groups having at least oneheteroatom in at least one ring, and preferably 5 or 6 atoms in eachring. The heteroaryl group preferably has 1 or 2 oxygen atoms and/or 1to 4 nitrogen atoms in the ring, and is bonded to the remainder of themolecule through a carbon. Exemplary heteroaryls include furyl,benzofuryl, oxazolyl, isoxazolyl, oxadiazolyl, benzoxazolyl,benzoxadiazolyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl,pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, indolyl, isoindolyl,indolizinyl, benzimidazolyl, indazolyl, benzotriazolyl,tetrazolopyridazinyl, carbazolyl, purinyl, quinolinyl, isoquinolinyl,imidazopyridyl and the like. Exemplary substituents include one or moreof the following groups: hydrocarbyl, substituted hydrocarbyl, hydroxy,protected hydroxy, acyl, acyloxy, alkoxy, alkenoxy, alkynoxy, aryloxy,halogen, amido, amino, cyano, ketals, acetals, esters and ethers.

The terms “hydrocarbon” and “hydrocarbyl” as used herein describeorganic compounds or groups consisting exclusively of the elementscarbon and hydrogen. These moieties include alkyl, alkenyl, alkynyl,aryl, carbocyclic moieties, and any combination of two or more thereof.These moieties also include alkyl, alkenyl, alkynyl, and aryl moietiessubstituted with other aliphatic or cyclic hydrocarbon groups, such asalkaryl, alkenaryl and alkynaryl. Unless otherwise indicated, thesemoieties preferably comprise 1 to 30 total carbon atoms.

As used herein, the term “organic” and grammatical variations thereof,in reference to a group or moiety, refers to a material comprisingcarbon, typically in combination with at least some hydrogen, andoptionally including one or more other elements, such as oxygen, sulfur,nitrogen, phosphorous, a halogen, or another non-metal or metalloidelement from groups II-A (e.g., B), IV-A (e.g., Si), V-A (e.g., As),VI-A (e.g., Se) of the Periodic Table. The term “organic” also refers tomaterials traditionally described as organometallic materials (e.g.,comprising one or more main group of or transition metal atomscovalently bound to a carbon atom), as well as materials that includemetallic elements in a complex or as a salt with an organic moiety.Non-limiting examples of organic moieties or groups include,hydrocarbons, heterocycles (including materials comprising at least onesaturated, unsaturated and/or aromatic ring comprising at least onecarbon atom, and one or more other elements), carbohydrates (includingsugars and polysaccharides), amino acids, polypeptides (includingproteins and other materials comprising at least two amino acid groupsbound together via a peptide bond), peptide analogs (including materialscomprising two or more amino acids linked by a bond other than a peptidebond, e.g., ester bonds), and a combination of two or more thereof.

The “substituted” moieties described herein (e.g., substitutedhydrocarbyl, heteroaryl, aryl and heterocyclic moieties) are groups thatare substituted with a group comprising at least one atom other thancarbon, including moieties in which a carbon chain atom is substitutedwith a hetero atom such as nitrogen, oxygen, silicon, phosphorous,boron, sulfur, or a halogen atom. In some embodiments, thesesubstituents include, e.g., one or more of halogen (F, Cl, Br, I),heterocyclo, alkoxy, alkenoxy, alkynoxy, aryloxy, hydroxy, protectedhydroxy, acyl, acyloxy, nitro, amino, amido, nitro, cyano, ketals,acetals, esters and ethers. In some embodiments such substituent groupscan be, e.g., —OR⁴, —N(R⁴)₂, —N⁺(R⁴)₃, —SR⁴, —OC(═O)R⁴, —N(R⁴)C(═O)R⁴,—SC(═O)R⁴, —OC(═O)OR⁵, —N(R⁴)C(═O)OR⁵, —SC(═O)OR⁵, —OC(═O)N(R⁴)₂,—N(R⁴)C(═O)N(R⁴)₂, —SC(═O)N(R⁴)₂, —OC(═O)SR⁵, —N(R⁴)C(═O)SR⁵,—SC(═O)SR⁵, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)N(R⁴)₂, —C(═O)SR⁴, —OC(═NR⁴)R⁴,—N(R⁴)C(═NR⁴)R⁴, —SO₂R⁴, —SO₂OR⁴, —SO₂(NR⁴)₂, —N(R⁴)SO₂OR⁵,—N(R⁴)SO₂N(R⁴)₂, —OSO₂OR⁵, —OSO₂N(R⁴)₂, —P(═O)(OR⁴)₂, —OP(═O)(OR⁴)₂,—OP(═O)R⁵(OR⁴), fluoro, chloro, bromo, iodo, —NO₂, —N₃, —N═N—Ar¹, —CN, aheteroaryl moiety, or a nonaromatic heterocyclic moiety; wherein each R⁴independently is H, hydrocarbyl, heteroaryl, or a nonaromaticheterocyclic moiety; each R⁵ independently is hydrocarbyl, heteroaryl,or a nonaromatic heterocyclic moiety; and each Ar¹ independently is arylor heteroaryl, which can be substituted as described above, or can beunsubstituted.

The use of the terms “a” and “an” and “the” and similar referents in thecontext of describing the invention (especially in the context of thefollowing claims) are to be construed to cover both the singular and theplural, unless otherwise indicated herein or clearly contradicted bycontext. The terms “comprising,” “having,” “including,” and “containing”are to be construed as open-ended terms (i.e., meaning “including, butnot limited to,”) unless otherwise noted. Recitation of ranges of valuesherein are merely intended to serve as a shorthand method of referringindividually to each separate value falling within the range, unlessotherwise indicated herein, and each separate value is incorporated intothe specification as if it were individually recited herein. Allnumerical values obtained by measurement (e.g., weight, concentration,physical dimensions, removal rates, flow rates, and the like) are not tobe construed as absolutely precise numbers, and should be considered toencompass values within the known limits of the measurement techniquescommonly used in the art, regardless of whether or not the term “about”is explicitly stated. All methods described herein can be performed inany suitable order unless otherwise indicated herein or otherwiseclearly contradicted by context. The use of any and all examples, orexemplary language (e.g., “such as”) provided herein, is intended merelyto better illuminate certain aspects of the invention and does not posea limitation on the scope of the invention unless otherwise claimed. Nolanguage in the specification should be construed as indicating anynon-claimed element as essential to the practice of the invention.

“Pharmaceutically or pharmacologically acceptable” include molecularentities and compositions that do not produce an adverse, allergic orother untoward reaction when administered to an animal, or a human, asappropriate. For human administration, preparations should meetsterility, pyrogenicity, and general safety and purity standards asrequired by FDA Office of Biologics standards.

The term “pharmaceutically acceptable carrier” or “pharmaceuticallyacceptable excipient” as used herein refers to any and all solvents,dispersion media, coatings, isotonic and absorption delaying agents, andthe like, that are compatible with pharmaceutical administration. Theuse of such media and agents for pharmaceutically active substances iswell known in the art. The compositions may also contain other activecompounds providing supplemental, additional, or enhanced therapeuticfunctions.

The term “pharmaceutical composition” as used herein refers to acomposition comprising at least one compound as disclosed hereinformulated together with one or more pharmaceutically acceptablecarriers, vehicles, or diluents.

“Individual,” “patient,” or “subject” are used interchangeably andinclude any animal, including mammals, preferably mice, rats, otherrodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates,and most preferably humans. Disclosed compounds may be administered to amammal, such as a human, but may also be administered to other mammalssuch as an animal in need of veterinary treatment, e.g., domesticanimals (e.g., dogs, cats, and the like), farm animals (e.g., cows,sheep, pigs, horses, and the like) and laboratory animals (e.g., rats,mice, guinea pigs, and the like). “Modulation” includes antagonism(e.g., inhibition), agonism, partial antagonism and/or partial agonism.

In the present specification, the term “therapeutically effectiveamount” means the amount of the subject compound that will elicit thebiological or medical response of a tissue, system or animal, (e.g.mammal or human) that is being sought by the researcher, veterinarian,medical doctor or other clinician. The compounds of the invention areadministered in therapeutically effective amounts to treat a disease.Alternatively, a therapeutically effective amount of a compound is thequantity required to achieve a desired therapeutic and/or prophylacticeffect.

The term “pharmaceutically acceptable salt(s)” as used herein refers tosalts of acidic or basic groups that may be present in compounds used inthe compositions. Compounds included in the present compositions thatare basic in nature are capable of forming a wide variety of salts withvarious inorganic and organic acids. The acids that may be used toprepare pharmaceutically acceptable acid addition salts of such basiccompounds are those that form non-toxic acid addition salts, i.e., saltscontaining pharmacologically acceptable anions, including, but notlimited to, malate, oxalate, chloride, bromide, iodide, nitrate,sulfate, bisulfate, phosphate, acid phosphate, isonicotinate, acetate,lactate, salicylate, citrate, tartrate, oleate, tannate, pantothenate,bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate,gluconate, glucaronate, saccharate, formate, benzoate, glutamate,methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonateand pamoate (i.e., 1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts.Compounds included in the present compositions that are acidic in natureare capable of forming base salts with various pharmacologicallyacceptable cations. Examples of such salts include alkali metal oralkaline earth metal salts, particularly calcium, magnesium, sodium,lithium, zinc, potassium, and iron salts. Compounds included in thepresent compositions that include a basic or acidic moiety may also formpharmaceutically acceptable salts with various amino acids. Thecompounds of the disclosure may contain both acidic and basic groups;for example, one amino and one carboxylic acid group. In such a case,the compound can exist as an acid addition salt, a zwitterion, or a basesalt.

The compounds disclosed herein contain one or more chiral centers and,therefore, exist as stereoisomers. The term “stereoisomers” when usedherein consists of all enantiomers or diastereomers. These compounds maybe designated by the symbols “(+),” “(−),” “R” or “S,” depending on theconfiguration of substituents around the stereogenic carbon atom, butthe skilled artisan will recognize that a structure may denote a chiralcenter implicitly. The present invention encompasses variousstereoisomers of these compounds and mixtures thereof. Mixtures ofenantiomers or diastereomers may be designated “(±)” in nomenclature,but the skilled artisan will recognize that a structure may denote achiral center implicitly.

The compounds disclosed herein may contain one or more double bonds and,therefore, exist as geometric isomers resulting from the arrangement ofsubstituents around a carbon-carbon double bond. The symbol

denotes a bond that may be a single, double or triple bond as describedherein. Substituents around a carbon-carbon double bond are designatedas being in the “Z” or “E” configuration wherein the terms “Z” and “E”are used in accordance with IUPAC standards. Unless otherwise specified,structures depicting double bonds encompass both the “E” and “Z”isomers. Substituents around a carbon-carbon double bond alternativelycan be referred to as “cis” or “trans,” where “cis” representssubstituents on the same side of the double bond and “trans” representssubstituents on opposite sides of the double bond.

Compounds disclosed herein may contain a carbocyclic or heterocyclicring and therefore, exist as geometric isomers resulting from thearrangement of substituents around the ring. Substituents around acarbocyclic or heterocyclic ring may be referred to as “cis” or “trans”,where the term “cis” represents substituents on the same side of theplane of the ring and the term “trans” represents substituents onopposite sides of the plane of the ring.

Mixtures of compounds wherein the substituents are disposed on both thesame and opposite sides of plane of the ring are designated “cis/trans.”

Individual enantiomers and diastereomers of contemplated compounds canbe prepared synthetically from commercially available starting materialsthat contain asymmetric or stereogenic centers, or by preparation ofracemic mixtures followed by resolution methods well known to those ofordinary skill in the art. These methods of resolution are exemplifiedby (1) attachment of a mixture of enantiomers to a chiral auxiliary,separation of the resulting mixture of diastereomers byrecrystallization or chromatography and liberation of the optically pureproduct from the auxiliary, (2) salt formation employing an opticallyactive resolving agent, (3) direct separation of the mixture of opticalenantiomers on chiral liquid chromatographic columns or (4) kineticresolution using stereoselective chemical or enzymatic reagents. Racemicmixtures can also be resolved into their component enantiomers bywell-known methods, such as chiral-phase liquid chromatography orcrystallizing the compound in a chiral solvent. Stereoselectivesyntheses, a chemical or enzymatic reaction in which a single reactantforms an unequal mixture of stereoisomers during the creation of a newstereocenter or during the transformation of a pre-existing one, arewell known in the art. Stereoselective syntheses encompass both enantio-and diastereoselective transformations, and may involve the use ofchiral auxiliaries. For examples, see Carreira and Kvaerno, Classics inStereoselective Synthesis, Wiley-VCH: Weinheim, 2009.

The compounds disclosed herein can exist in solvated as well asunsolvated forms with pharmaceutically acceptable solvents such aswater, ethanol, and the like. It is intended that the invention embraceboth solvated and unsolvated forms. In one embodiment, the compound isamorphous. In one embodiment, the compound is a single polymorph. Inanother embodiment, the compound is a mixture of polymorphs. In anotherembodiment, the compound is in a crystalline form.

The invention also embraces isotopically labeled compounds as disclosedherein which are identical to those recited herein, except that one ormore atoms are replaced by an atom having an atomic mass or mass numberdifferent from the atomic mass or mass number usually found in nature.Examples of isotopes that can be incorporated into compounds of theinvention include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorus, sulfur, fluorine and chlorine, such as ²H, ³H, ¹³C, 14C,¹⁵N, ¹⁸O, ¹⁷O ³¹P, ³²P, ³⁵S ¹⁸F, and ³⁶Cl, respectively. For example, acompound of the invention may have one or more H atoms replaced withdeuterium.

Certain isotopically-labeled disclosed compounds (e.g., those labeledwith ³H and ¹⁴C) are useful in compound and/or substrate tissuedistribution assays. Tritiated (i.e., ³H) and carbon-14 (i.e., ¹⁴C)isotopes are particularly preferred for their ease of preparation anddetectability. Further, substitution with heavier isotopes such asdeuterium (i.e., ²H) may afford certain therapeutic advantages resultingfrom greater metabolic stability (e.g., increased in vivo half-life orreduced dosage requirements) and hence may be preferred in somecircumstances. Isotopically labeled compounds of the invention cangenerally be prepared by following procedures analogous to thosedisclosed in the examples herein by substituting an isotopically labeledreagent for a non-isotopically labeled reagent.

Compounds and pharmaceutical compositions (e.g., therapeutic agents ormedicaments, and compositions comprising the compounds or therapeuticagents or medicaments) are described herein, which comprise a compoundrepresented by Formula (I):

wherein: Y is a biologically active organic core group comprising one ormore unsubstituted or substituted moiety selected from an aryl group, aheteroaryl aryl group, a nonaromatic hydrocarbyl group, and anonaromatic heterocyclic group, to which each Z independently iscovalently bonded; n is 1, 2, 3, 4 or 5; each Z independently is O, NR,or N; when Z is O, m is 1, X¹ is a covalent bond, and the Z iscovalently bonded to an aryl or heteroaryl moiety of Y; when Z is NR, mis 1, X¹ is a covalent bond or CH₂CH₂, and the Z is covalently bonded toa nonaromatic hydrocarbyl, a nonaromatic heterocyclic, an aryl, orheteroaryl moiety of Y; when Z is N, either (a) m is 2, X¹ is CH₂CH₂,and the Z is covalently bonded to a nonaromatic hydrocarbyl, anonaromatic heterocyclic, an aryl, or a heteroaryl moiety of Y; or (b) mis 1, X¹ is a covalent bond or CH₂CH₂, and the Z is a nitrogen in anaromatic or non-aromatic heterocyclic ring portion of core group Y; eachX² independently is O or NR; and each R independently comprises H or asubstituted or unsubstituted group selected from an aryl group, aheteroaryl aryl group, a nonaromatic hydrocarbyl group, and anonaromatic heterocyclic group.

In some embodiments, a therapeutic compound or medicament represented byFormula (II) is described:

wherein A is an organic moiety comprising at least one substituent, R¹;n is 1, 2, 3, 4 or 5; each Z independently is O, NR, or N; each Z iscovalently bonded to an R^(i) moiety of A; each R independentlycomprises a hydrocarbyl group; when Z is O, m is 1, and X¹ is a covalentbond. Each X² independently can be O or NR⁵ (preferably, X² is O). WhenZ is NR, m is 1, and each X¹ independently is a covalent bond or CH₂CH₂.When Z is N, m is 2, and X¹ is CH₂CH₂. Each R¹ independently is an arylgroup, a heteroaryl group, and a substituted aryl group having theformula:

Each R² and R³ independently is a substituent selected from the groupconsisting of a hydrocarbyl moiety, —OR⁴, —N(R⁴)₂, —N⁺(R⁴)₃, —SR⁴,—OC(═O)R⁴, —N(R⁴)C(═O)R⁴, —SC(═O)R⁴, —OC(═O)OR⁵, —N(R⁴)C(═O)OR⁵,—SC(═O)OR⁵, —OC(═O)N(R⁴)₂, —N(R⁴)C(═O)N(R⁴)₂, —SC(═O)N(R⁴)₂, —OC(═O)SR⁵,—N(R⁴)C(═O)SR⁵, —SC(═O)SR⁵, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)N(R⁴)₂,—C(═O)SR⁴, —OC(═NR⁴)R⁴, —N(R⁴)C(═NR⁴)R⁴, —SO₂R⁴, —SO₂OR⁴, —SO₂(NR⁴)₂,—N(R⁴)SO₂OR⁵, —N(R⁴)SO₂N(R⁴)₂, —OSO₂OR⁵, —OSO₂N(R⁴)₂, —P(═O)(OR⁴)₂,—OP(═O)(OR⁴)₂, —OP(═O)R⁵(OR⁴), fluoro, chloro, bromo, iodo, —NO₂, —N₃,—N═N—Ar¹, —CN, a heteroaryl moiety, and a nonaromatic heterocyclicmoiety. Alternatively, an R² and an R³ together form a ring selectedfrom a fused 5-member nonaromatic carbocyclic ring, a fused 5-memberheterocyclic ring, a fused 6-member nonaromatic carbocyclic ring, and afused 6-member nonaromatic nitrogen-containing heterocyclic ring. EachR⁴ independently is H, hydrocarbyl, heteroaryl, or a nonaromaticheterocyclic moiety. Each R⁵ independently is hydrocarbyl, heteroaryl,or a nonaromatic heterocyclic moiety. Each Ar¹ independently is aryl orheteroaryl. Each Ar¹, hydrocarbyl, heteroaryl, nonaromatic heterocyclicmoiety, fused 5-member nonaromatic carbocyclic ring, fused 5-memberheterocyclic ring, fused 6-member nonaromatic carbocyclic ring, andfused 6-member nonaromatic nitrogen-containing heterocyclic ringindependently is unsubstituted or is substituted with a group comprisingat least one atom other than carbon. The parameters x and y are 0, 1 or2; and the sum of x and y is at least 1 when R¹ is the substituted aryl.

Certain compounds of Formula (II) or Formula (I) can be made by reactinga compound of Formula (IV) or (V), respectively, with SO₂F₂ in thepresence of a base:

in which A, Y, m and n are as defined for compounds of Formulas (II) and(I), and Z is O or NR. Examples of suitable bases include (e.g., analkali metal hydroxide, such as NaOH, KOH and the like), an alkali metalalkoxide (e.g., potassium tert-butoxide, sodium methoxide, and thelike), a nitrogen base (preferably a tertiary amine, such astriethylamine or diisopropylethylamine; an amidine such as DBU; aguanidine such as tetramethylguanidine), and the like.

In another embodiment, a method of preparing a compound of Formula (II)or Formula (I) in which each m is 2, each Z is N, and each X¹ is CH₂CH₂,comprises reacting a compound of Formula (IV) or (V), respectively, withCH₂═CH—SO₂F (“ESF”), which readily condenses with amino compoundscomprising at least one N—H bond.

The incorporation of a fluorosulfonyl (e.g., as CH₂CH₂SO₂F) orfluorosulfonyloxy (i.e., —OSO₂F) group into a therapeutically activecompound (a medicament) in many cases surprisingly increases themetabolic stability of such compounds and contributes tobioavailability. In some cases, a non-covalent drug can be converted toa covalent drug by the incorporation of —SO₂F or —OSO₂F. In other cases,the solubility of the compounds of Formula (II) are enhanced relative tothe parent therapeutic agents comprising the core, for example when an—OSO₂F replaces a CF₃ or OCF₃ group.

The therapeutically active compounds can be those which target apathogen, as well as compounds that target a site of action in a hostsubject (e.g., a patient).

Illustrative therapeutically active compounds that target a pathogen andare suitable for incorporation of a —SO₂F or —OSO₂F group include, e.g.,antibiotics such as vancomycin, rifamycin, rifampicin, teicoplanin,sulfacetamide, amoxicillin, novobiocin, tetracyclines (e.g.,tetracycline, oxytetracycline, methacycline, minocycline,chlorotetracycline, doxycycline, rolitetracycline, demeclocycline, andthe like), sulfanilamide, sulfamethoxazole, norfloxacin, gatifloxacin,gemifloxacin, trimethoprim, pyrimethamine, cefadroxil, anti-tubercularantibiotics (e.g., isoniazid, rifampicin; streptomycin, ciprofloxacin,moxifloxacin, aminosalicylic acid, and the like); and anti-protozoalagents such as iodoquinol, paramomycin, anti-malarial agents (e.g.,quinine (by replacement of OMe with OSO₂F), quinocrine, atovaquone(e.g., by replacement of Cl or by reaction with an OH), mefloquine,sulfadoxine, hydrochloroquine, proguanil (e.g., by replacement of Clwith OSO₂F), and the like.

Illustrative therapeutically active compounds that target a site ofactivity in host subject and are suitable for incorporation of a —SO₂For —OSO₂F group include, e.g., non-steroidal anti-inflammatory agents(NSAIDs) such as naproxen, ibuprofen, aspirin, tolmetin, flurbiprofen,sulindac, piroxicam, nabumeton, flufenamic acid, tolfenamic acid,diclofenac, and the like; antineoplastic agents such as bleomycin,cytarabine, dacarbazine, anthracyclines (e.g., daunorubicin,doxorubicin, and the like), epirubicin, etoposide, flutamide,gemcitabine, idarubicin, leuprolide, leuprorelin, mercaptopurine,methotrexate, mitomycin, mitoxantrone, pemetrexed, pentostatin,procarbazine, suramin, teniposide, thioguanine, thiotepa, uracil mustard(uramastine), and the like; opiates such as morphine, buprenorphine,hydromorphone, oxymorphone, dihydromorphone, methyldihydromorphinone,butorphanol, and the like; analgesics such as pregabalin,tetrahydrocannabinol, fentanyl, flupirtine, oxycodone, acetaminophen,salicylamide, and the like; anti-depressants such as fluoxetine(PROZAC), sertraline (ZOLOFT), duloxetine (CYMBALTA), amoxapine,maprotiline, mianserin, nomifensin, trazodine, viloxazine, aripirazole,bupropion (WELLBUTRIN), desvenlafaxine, duloxetine, paroxetine, and thelike; COX 2 inhibitors such as celecoxib, rofecoxib, lumiracoxib,etoricoxib, firocoxib, nimesulide, and the like; COX-LOX inhibitors suchas licofelone, clonidine, and the like; opioid receptor antagonists suchas naltrexone, naloxone, naltrindole, and the like; Alzheimer's diseasemedications such as epigallocatechin gallate (EGCG), memantine,galantamine, and the like; statins such as atorvstatin (LIPITOR),rosuvastatin, and the like; erectile dysfunction medications such assildenafil (VIAGRA), tadalafil (CIALIS), vardenafil (LEVITRA),apomorphine, and the like; anti-asthma medications such as salbutamol(albuterol), salmeterol, terbutaline, formoterol, metaproterenol, andthe like; cholinesterase inhibitors such as edrophonium, tacrine, andthe like; sympathomimetic drugs such as phenylephrine, amphetamine,methoxamine, prenalterol, terbutaline, ritodrine, and the like;anti-seizure agents such as lamotrigine, vigabatrine, gabapentin,pregabalin, and the like; neuromuscular blockers such as tubocurarine,cisatracurium, and the like; intestinal steroid absorption inhibitorssuch as ezetimibe,(3R,4S)-1,4-bis(4-methoxyphenyl)-3-(3-phenylpropyl)-2-azetidinone, andthe like; endocrine drugs such as thyroxine, somatostatin, and the like;estrogenic agents, antagonists and agonists, such as raloxifene,estradiol, ethynylestradiol, diethylstilbesrol, and the like; antiviralagents such as acyclovir, valacyclovir, penciclovir, cidofovir,zalcitibine, adefovir, entacavir, and the like; anorectic agents such asphentermine, and the like; anticoagulants such as warfarin,acenocoumarol, and the like; antihypertives and beta blockers such aslisinopril, nadolol, atenolol, acebutolol, betaxolol, carvediol,esmolol, and the like; seratonin receptor agonists and seratonin uptakeinhibitors such as seratonin, sertraline, dolasetron, fluoxetine, andthe like; diuretics such as hydrochlorothiazide, bumetanide, furosemide,pinoresinol, and the like; calcium channel blockers such as amlodipinebesylate, mibefradin hydrochloride, and the like; as well as femalelibido enhancing compounds such as flibanserin(1-(2-{4-[3-(Trifluoromethyl)phenyl]piperazin-1-yl}ethyl)-1,3-dihydro-2H-benzimidazol-2-one;Sprout Pharmaceuticals). Other suitable materials include peptide-basedand amino acid-based agents, particularly tyrosine,2,6-dimethyltyrosine, lysine, and peptides comprising one or moreresidues selected from tyrosine, 2,6-dimethyltyrosine, and lysine suchas leuprolide (ENANTONE, a tyrosine-containing peptide pituitary GnRHreceptor antagonist), glatiramer (a random copolymer of lysine alanineaspartic acid and tyrosine, tradename CAPOXONE, an immunomodulator), andthe like. As is well known in the medical art, drugs within in aparticular classification (e.g., antibiotic, estrogenic agent,antineoplastic agent, and the like) may have therapeutic uses andindications for more than one type of disease or condition.

In many cases, the SO₂F group can attach to the biologically active coreby replacement of a hydrogen of an aromatic or heteroaromatic OH or ahydrogen of an amino group of the core to form the an —OSO₂F, or —NRSO₂Fgroup. Particularly in the case of amino groups bearing a hydrogen atompresent in the medicament structure, a NRCH₂CH₂SO₂F or N(CH₂CH₂SO₂F)₂group can be introduced by replacement of the hydrogen atom. Thesereplacements are readily accomplished by reaction of virtually any OH orNHR group with SO₂F₂ in the former case and reaction of an NH₂ or NHRwith ESF in the latter cases. In other embodiments, an OSO₂F group canbe attached to the medicament as a replacement for a methoxy ortrifluoromethoxy group, or can be added to the medicament in place of ahydrogen of a CH or in place of some other substituent group by methodsof organic synthesis that are well known in the chemical arts.Preferably, the SO₂F group is attached to the medicament by replacementof a hydrogen of an aromatic or heteroaromatic OH or a hydrogen of anamino group as described herein. Compounds comprising a —S(O)(NR⁵)Fgroup can be obtained by replacement of Cl from a corresponding—S(O)(NR⁵)Cl prepared by well-known conventional means as describedherein.

Therapeutically active compounds comprising —S(O)(X²)F groups (e.g.,NCH₂CH₂SO₂F, NSO₂F, OSO₂F and/or S(O)(NR⁵)F groups) described herein aresuitable, e.g., as medicaments for humans and animals, since thesefunctional groups generally do not significantly interfere with thebiological/therapeutic activity of the parent therapeutic agents. Inaddition, the —S(O)(X²)F groups provide useful handles for selectivelyderivatizing the therapeutic agent, e.g., to add a useful functional ordiagnostic group such as a dye, biotin, and the like.

Such therapeutic compounds can be formulated as a pharmaceuticalcomposition in combination with a pharmaceutically acceptable carrier,vehicle, or diluent, such as an aqueous buffer at a physiologicallyacceptable pH (e.g., pH 7 to 8.5), a polymer-based nanoparticle vehicle,a liposome, and the like. The pharmaceutical compositions can bedelivered in any suitable dosage form, such as a liquid, gel, solid,cream, or paste dosage form. In one embodiment, the compositions can beadapted to give sustained release of the compound of Formula (I).

Pharmaceutical compositions comprising therapeutic compounds of Formula(I) can be administered to a subject or patient in a therapeuticallyeffective amount to treat a disease or condition, e.g., a disease orcondition for which the biologically active core group, A, is active.

In some embodiments, the pharmaceutical compositions include, but arenot limited to, those forms suitable for oral, rectal, nasal, topical,(including buccal and sublingual), transdermal, vaginal, or parenteral(including intramuscular, subcutaneous, and intravenous) administration,in a form suitable for administration by inhalation or insufflation, orinjection into amniotic fluid. The compositions can, where appropriate,be conveniently provided in discrete dosage units. The pharmaceuticalcompositions of the invention can be prepared by any of the methods wellknown in the pharmaceutical arts. Some preferred modes of administrationinclude intravenous (iv), topical, subcutaneous, and injection intoamniotic fluid.

Pharmaceutical formulations suitable for oral administration includecapsules, cachets, or tablets, each containing a predetermined amount ofone or more of the compounds of Formula (I), as a powder or granules. Inanother embodiment, the oral composition is a solution, a suspension, oran emulsion. Alternatively, the compounds of Formula (I) can be providedas a bolus, electuary, or paste. Tablets and capsules for oraladministration can contain conventional excipients such as bindingagents, fillers, lubricants, disintegrants, colorants, flavoring agents,preservatives, or wetting agents. The tablets can be coated according tomethods well known in the art, if desired. Oral liquid preparationsinclude, for example, aqueous or oily suspensions, solutions, emulsions,syrups, or elixirs. Alternatively, the compositions can be provided as adry product for constitution with water or another suitable vehiclebefore use. Such liquid preparations can contain conventional additivessuch as suspending agents, emulsifying agents, non-aqueous vehicles(which may include edible oils), preservatives, and the like. Theadditives, excipients, and the like typically will be included in thecompositions for oral administration within a range of concentrationssuitable for their intended use or function in the composition, andwhich are well known in the pharmaceutical formulation art. Thecompounds of Formula (I) will be included in the compositions within atherapeutically useful and effective concentration range, as determinedby routine methods that are well known in the medical and pharmaceuticalarts. For example, a typical composition can include one or more of thecompounds of Formula (I) at a concentration in the range of at leastabout 0.01 nanomolar to about 1 molar, preferably at least about 1nanomolar to about 100 millimolar.

Pharmaceutical compositions for parenteral administration (e.g. by bolusinjection or continuous infusion) or injection into amniotic fluid canbe provided in unit dose form in ampoules, pre-filled syringes, smallvolume infusion, or in multi-dose containers, and preferably include anadded preservative. The compositions for parenteral administration canbe suspensions, solutions, or emulsions, and can contain excipients suchas suspending agents, stabilizing agent, and dispersing agents.Alternatively, the compounds of Formula (I) can be provided in powderform, obtained by aseptic isolation of sterile solid or bylyophilization from solution, for constitution with a suitable vehicle,e.g. sterile, pyrogen-free water, before use. The additives, excipients,and the like typically will be included in the compositions forparenteral administration within a range of concentrations suitable fortheir intended use or function in the composition, and which are wellknown in the pharmaceutical formulation art. The compounds of Formula(I) will be included in the compositions within a therapeutically usefuland effective concentration range, as determined by routine methods thatare well known in the medical and pharmaceutical arts. For example, atypical composition can include one or more of the compounds of Formula(I) at a concentration in the range of at least about 0.01 nanomolar toabout 100 millimolar, preferably at least about 1 nanomolar to about 10millimolar.

Pharmaceutical compositions for topical administration of the compoundsto the epidermis (mucosal or cutaneous surfaces) can be formulated asointments, creams, lotions, gels, or as a transdermal patch. Suchtransdermal patches can contain penetration enhancers such as linalool,carvacrol, thymol, citral, menthol, t-anethole, and the like. Ointmentsand creams can, for example, include an aqueous or oily base with theaddition of suitable thickening agents, gelling agents, colorants, andthe like. Lotions and creams can include an aqueous or oily base andtypically also contain one or more emulsifying agents, stabilizingagents, dispersing agents, suspending agents, thickening agents,coloring agents, and the like. Gels preferably include an aqueouscarrier base and include a gelling agent such as cross-linkedpolyacrylic acid polymer, a derivatized polysaccharide (e.g.,carboxymethyl cellulose), and the like. The additives, excipients, andthe like typically will be included in the compositions for topicaladministration to the epidermis within a range of concentrationssuitable for their intended use or function in the composition, andwhich are well known in the pharmaceutical formulation art. Thecompounds of Formula (I) will be included in the compositions within atherapeutically useful and effective concentration range, as determinedby routine methods that are well known in the medical and pharmaceuticalarts. For example, a typical composition can include one or more of thecompounds of Formula (I) at a concentration in the range of at leastabout 0.01 nanomolar to about 1 molar, preferably at least about 1nanomolar to about 100 millimolar.

Pharmaceutical compositions suitable for topical administration in themouth (e.g., buccal or sublingual administration) include lozengescomprising the compound in a flavored base, such as sucrose, acacia, ortragacanth; pastilles comprising the peptide in an inert base such asgelatin and glycerin or sucrose and acacia; and mouthwashes comprisingthe active ingredient in a suitable liquid carrier. The pharmaceuticalcompositions for topical administration in the mouth can includepenetration enhancing agents, if desired. The additives, excipients, andthe like typically will be included in the compositions of topical oraladministration within a range of concentrations suitable for theirintended use or function in the composition, and which are well known inthe pharmaceutical formulation art. The compounds of Formula (I) will beincluded in the compositions within a therapeutically useful andeffective concentration range, as determined by routine methods that arewell known in the medical and pharmaceutical arts. For example, atypical composition can include one or more of the compounds of Formula(I) at a concentration in the range of at least about 0.01 nanomolar toabout 1 molar, preferably at least about 1 nanomolar to about 100millimolar.

A pharmaceutical composition suitable for rectal administrationcomprises a compound of the present invention in combination with asolid or semisolid (e.g., cream or paste) carrier or vehicle. Forexample, such rectal compositions can be provided as unit dosesuppositories. Suitable carriers or vehicles include cocoa butter andother materials commonly used in the art. The additives, excipients, andthe like typically will be included in the compositions of rectaladministration within a range of concentrations suitable for theirintended use or function in the composition, and which are well known inthe pharmaceutical formulation art. The compounds of Formula (I) will beincluded in the compositions within a therapeutically useful andeffective concentration range, as determined by routine methods that arewell known in the medical and pharmaceutical arts. For example, atypical composition can include one or more of the compounds of Formula(I) at a concentration in the range of at least about 0.01 nanomolar toabout 1 molar, preferably at least about 1 nanomolar to about 100millimolar.

According to one embodiment, pharmaceutical compositions of the presentinvention suitable for vaginal administration are provided as pessaries,tampons, creams, gels, pastes, foams, or sprays containing a compound ofFormula (I) of the invention in combination with a carriers as are knownin the art. Alternatively, compositions suitable for vaginaladministration can be delivered in a liquid or solid dosage form. Theadditives, excipients, and the like typically will be included in thecompositions of vaginal administration within a range of concentrationssuitable for their intended use or function in the composition, andwhich are well known in the pharmaceutical formulation art. Thecompounds of Formula (I) will be included in the compositions within atherapeutically useful and effective concentration range, as determinedby routine methods that are well known in the medical and pharmaceuticalarts. For example, a typical composition can include one or more of thecompounds of Formula (I) at a concentration in the range of at leastabout 0.01 nanomolar to about 1 molar, preferably at least about 1nanomolar to about 100 millimolar.

Pharmaceutical compositions suitable for intra-nasal administration arealso encompassed by the present invention. Such intra-nasal compositionscomprise a compound of Formula (I) in a vehicle and suitableadministration device to deliver a liquid spray, dispersible powder, ordrops. Drops may be formulated with an aqueous or non-aqueous base alsocomprising one or more dispersing agents, solubilizing agents, orsuspending agents. Liquid sprays are conveniently delivered from apressurized pack, an insufflator, a nebulizer, or other convenient meansof delivering an aerosol comprising the peptide. Pressurized packscomprise a suitable propellant such as dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, orother suitable gas as is well known in the art. Aerosol dosages can becontrolled by providing a valve to deliver a metered amount of thepeptide. Alternatively, pharmaceutical compositions for administrationby inhalation or insufflation can be provided in the form of a drypowder composition, for example, a powder mix of the compounds ofFormula (I) and a suitable powder base such as lactose or starch. Suchpowder composition can be provided in unit dosage form, for example, incapsules, cartridges, gelatin packs, or blister packs, from which thepowder can be administered with the aid of an inhalator or insufflator.The additives, excipients, and the like typically will be included inthe compositions of intra-nasal administration within a range ofconcentrations suitable for their intended use or function in thecomposition, and which are well known in the pharmaceutical formulationart. The compounds of Formula (I) will be included in the compositionswithin a therapeutically useful and effective concentration range, asdetermined by routine methods that are well known in the medical andpharmaceutical arts. For example, a typical composition can include oneor more of the compounds of Formula (I) at a concentration in the rangeof at least about 0.01 nanomolar to about 1 molar, preferably at leastabout 1 nanomolar to about 100 millimolar.

Optionally, the pharmaceutical compositions of the present invention caninclude one or more other therapeutic agent, e.g., as a combinationtherapy.

Sulfonyl chlorides are the most commonly used S(VI) electrophiles.RSO₂Cl and ClSO₂Cl often cannot serve as reliable connective unitsbecause of the facile reductive failure of the bond between sulfur(VI)and chlorine (eq. 1). This emerges most vexingly in the attemptedformation of inorganic sulfate, sulfamide, and sulfamate linkages suchas RO—SO₂—OR′, RNH—SO₂—NHR′ and ArO—SO₂—NRR′. Attempts to develop quickand robust inorganic connectors for the fast assembly of sophisticatedmolecules has been delayed by these side reactions. As described herein,sulfonyl fluoride and related groups demonstrated to constitutecomponents of a versatile new click chemistry, encompassing both carbon-(C—SO₂F) and heteroatom-bound (N—SO₂F and O—SO₂F) species; see e.g.,Equation (1).

An understanding of the unique stability-reactivity pattern of sulfonylfluorides rests on five contributing factors, illustrated by the resultsin FIG. 1.

(1) Resistance to reduction. Since fluorine is the most electronegativeelement in the periodic table, sulfonyl-fluorine bond cleavage isexclusively heterolytic with the formation of fluoride ion (althoughrarely, if ever, as uncomplexed F). In contrast, homolytic scission ofS—Cl bonds is quite common. For aromatic cases, irreversible reductionto the sulfinic acid level (ArS(O)OR) occurs easily for ArSO₂Cl withmany nucleophiles, except alcohols and amines under limited conditions.Sulfonyl bromides and iodides are even more prone to reduction andradical reactions than sulfonyl chlorides, allowing sulfonyl chlorides,sulfonates, and sulfonic acids to be reduced cleanly if the iodide isgenerated in situ. A remarkable example of the differences betweenchlorine and fluorine in sulfuryl chemistry is provided by the parentcompounds: SO₂Cl₂ is a powerful oxidant, whereas sodium metal can bemelted in hot SO₂F₂ without chemical change in either species.

(2) Thermodynamic stability. Whereas substitution at all sulfur centersin oxidation states below VI is kinetically accessible, including thesulfur(IV) oxyhalides SOF₂ and SOCl₂, the very sluggishness of S(VI)substitution chemistry makes it superior as a connector. Furthermore,sulfonyl fluorides are much more stable than other sulfonyl halidestoward nucleophilic substitution (including hydrolysis) and thermolysis,making them the sulfonyl reagents of choice under demanding reactionconditions. These observations are consistent with the measured bondstrengths of SO₂—F relative to SO₂—Cl: the homolytic bond dissociationenergy of the S—F bond in SO₂F₂ (90.5±4.3 kcal/mol, 81±2 kcal/mol) isfar larger than the S—Cl bond in SO₂Cl₂ (46±4 kcal/mol). The differenceis of similarly large magnitude (41 kcal/mol) in comparing the bondstrengths of S—F vs. S—Cl bonds in SO₂FCl.

These factors produce a surprising and highly useful passivity in the—SO₂F group. An aliphatic example is provided by methanedisulfonylfluoride [(FSO₂)₂CH₂, MDSF]. The SO₂F groups in this compound survivesevere electrochemical oxidation conditions in the fluorination of themethylene group, and base-mediated and catalyzed alkylation andcondensation reactions proceed perfectly (Eq. 3). The chloride analogue(ClSO₂)₂CH₂ decomposes under these circumstances. As a building block,MDSF is especially useful for its dual potential to link withelectrophiles at carbon via its conjugate base and with nucleophiles ateach O₂S—F bond. Importantly, sulfonyl fluorides are surprisingly stableto aqueous conditions.

(3) Exclusive reaction at sulfur. Because of its polarizability, thechlorine center in —SO₂Cl and related groups is vulnerable tonucleophilic or reductive attack, so that reactions with carbonnucleophiles usually give mixtures of products resulting from bothsulfonylation and chlorination pathways.

(4) Special nature of the fluoride-proton interaction. Bothaddition-elimination and direct substitution pathways are reasonable fornucleophilic substitution reactions of sulfonyl fluorides. While thedetails of the SO₂ center's participation in this reaction are bothrelevant and incompletely understood, the key feature that makes SuFExchemistry unique is that it depends very much on stabilization of thedeveloping fluoride ion in the substitution process. Other halides canbe subject to similar effects, but fluoride stands alone in themagnitude and environmental sensitivity of the phenomenon. Furthermore,the agents that accomplish such fluoride stabilization under practicalconditions are H⁺ and silyl groups (FIG. 2), making the SuFEx processcontrollable and useful in both biological and synthetic settings.

The special nature of the fluoride ion in water has long beenrecognized, but is not often taken advantage of in a synthetic context.In FIG. 2, the role of “HX” in stabilizing fluoride represents thepotential virtues of specific protic centers in accelerating reactionsof —SO₂F electrophiles (as in reactions with proteins, discussed below),and the power of aqueous environments to transmit acidic stabilizationto the fluoride center. To understand the unique properties of fluorideas a base and leaving group, an appreciation of the bifluoride ion(HF₂)⁻ is essential. The bifluoride bond is a strong, centrosymmetric,three atom-four electron bond, worth a remarkable 40 kcal/mol. Thebifluoride bond is short and strong, and is not to be confused with thetime-honored, albeit weak, hydrogen bond/hydrogen bonding phenomenon.Hence, when fluoride encounters any acid in water, the bifluoride ion,[F—H—F]⁻, is formed, which also is in equilibrium with substantialquantities of higher adducts such as [F—H—F—H—F]⁻. In other words, F⁻ isa unique base: it gains strong stabilization in water by trapping aproton between two of itself. The proton is therefore uniquely effectiveat stabilizing fluoride as a leaving group. The reactivity of thebifluoride nucleophile, discussed below, is the complement to thehelpful role of hydrogen bonding in the chemistry of sulfonyl fluoridesin protic solvents.

(5) Closely Related Functional Groups.

Aliphatic sulfonylfluorides. Aryl sulfonyl fluorides are significantlymore resistant to hydrolysis than alkyl derivatives with

-hydrogens, and electron-withdrawing substituents on the aromatic ringincrease the electrophilic nature of S(VI) and make it more reactive.Sulfonyl halides, including fluorides, bearing acidic protons in the

-position undergo reactions that often proceed via elimination to formsulfene-type intermediates (RR′C═SO₂). A good example isphenylmethanesulfonyl fluoride (PMSF), a serine protease inhibitorwidely used in the preparation of cell lysates. However, this reactionpathway is fast only in the presence of base, allowing PMSF and otheraliphatic sulfonyl fluorides to be stable and to selectively modifyproteins in aqueous solution at moderate pH. Also noteworthy is the farbetter AlCl₃-assisted Friedel-Crafts reactivity of alkyl-SO₂F compoundscompared to alkyl-SO₂Cl. Thus, while the current focus is onarylsulfonyl connectors, aliphatic derivatives also benefit from theunique chemistry of the SO₂F group. The PMSF homologue with anadditional CH₂ group, i.e., PhCH₂CH₂SO₂F, is less reactive towardshydrolysis, whether enzymatic or otherwise.

Sulfonimidoylfluorides. Sulfonimidoyl fluorides generally have the sameadvantageous properties as sulfonyl fluorides, and reactivitycomparisons with sulfonimidoyl chlorides are similarly striking.However, the nitrogen substituent gives sulfonimidoyl fluorides anadditional point of modification, and their reactivities towardnucleophiles can be dramatically altered by the nature of thatsubstituent. Electron-withdrawing groups, such as acyl, carbonate andsulfonyl enhance the electrophilicity of S, making these classes ofcompounds similar in reactivity to sulfonyl fluorides, ranging from moreto less reactive. In contrast, sulfonimidoyl fluorides with alkyl andaryl groups on N are much more stable than sulfonyl chlorides, evenunder basic conditions (vide infra).

The most common processes of sulfonylation of aromatic and aliphaticmolecules are summarized in FIG. 3. The majority of these producesulfonyl chlorides, making them the least expensive and most availablesubstrates. The exchange of fluoride for chloride in these systems wouldseem to be a simple matter, but the transformation's history is unusualin several instructive ways.

The presence of water was found to be beneficial, and typical reactionconditions involve refluxing the water-organic biphasic mixtures.However, yields rarely exceed 80%. While “naked” fluoride (KF, dryacetonitrile, 18-crown-6) can be used, the bifluoride anion (F—H—F)⁻(e.g., from potassium bifluoride) is consistently and substantiallysuperior to other reagents for sulfonyl chloride-to-fluoride conversion,allowing the use of mild reaction conditions, broad substrate scope,simple reaction setup, effortless product isolation, and easy scale up.Bifluoride seems to be especially effective when it can be used “onwater”—that is, in reactions performed with a vigorously stirred oragitated water-organic interface. Since solvation and H-bonding isimportant to the state and reactivity of fluoride, [FHF]⁻ ataqueous-organic interfaces presents a more nucleophilic, albeit lessbasic and less solvated, fluoride source to electrophiles in the organicphase, shown in schematic fashion in FIG. 4. Strong acid (HX) has theeffect of enhancing the utility of fluoride as a nucleophile, but not asa base, by generating a form of the anion (bifluoride) that can bepresented more effectively at water interfaces.

Examples of sulfonyl fluorides made from the corresponding chlorides inthis way are shown in FIG. 5. If technical grade starting material isused, the sulfonyl fluoride product occasionally requires purificationby a wash with aqueous base and/or by chromatography on a short silicagel column. The crude product, however, is virtually free of impurities.Liquid sulfonyl chlorides are simply stirred vigorously with saturatedaqueous KFHF solution. Otherwise, acetonitrile (MeCN) generally is theco-solvent of choice. THF or CH₂Cl₂ optionally can be used as diluentsto dissolve a hydrophobic substrate and present it to the aqueousinterface where the reaction with bifluoride likely occurs. Fullconversion generally is achieved within several hours. When, as oftenhappens, starting chloride and product fluoride overlap on TLC, reactionprogress can be monitored by GC, LCMS, or ¹⁹F NMR.

Examples of the easy installation of alkyl (W. Qiu, D. J. Burton, J.Fluor. Chem. 1992, 60, 93-100) and aryl sulfonyl groups using thegeneral methods of FIG. 3 are shown in FIG. 6. In all cases, theintermediate sulfonyl chloride was subjected without purification to anaqueous phase of saturated KFHF. The desired fluoride product can beeasily purified if necessary by simple washing, recrystallization, orcolumn chromatography. Such in situ conversion to the fluoride isparticularly advantageous for certain heterocyclic sulfonyl chlorides,often generated by oxidation of thiols such as the 6-mercaptopurineshown in FIG. 6, that are unstable. KFHF, optimally already presentduring the Cl₂ oxidation stage, acts as both nucleophile and buffer, inthis case carrying the Het-SO₂—Cl on to the Het-SO₂—F before itcollapses to Het-Cl and SO₂.

Sulfonimidoyl chlorides and sulfamoyl chlorides with electronwithdrawing substituents on nitrogen are very similar in theirreactivity to sulfonyl chlorides (vide supra) and can be converted tothe corresponding fluorides by treatment with saturated aqueous KFHF(FIG. 7, Panels A,B). When electron donating groups are present onnitrogen, bifluoride is not reactive enough, giving low yields understandard conditions. In these cases, Bolm's silver fluoride inacetonitrile conditions are used to produce the sulfonyl fluoride on apreparative scale (FIG. 7, Panels C,D).

Several useful reagents are shown in FIG. 9, involving reactiveelectrophilic groups such as benzyl bromide, phenacyl bromide, acylhalide, isocyanate, and iodide. The lower reactivity of —SO₂F allowsthese reagents to be selectively attached via the other electrophilicsite. Azide- and alkyne-modified sulfonyl fluorides will also be usefulsince the SO₂F group does not interfere with any form of the catalyzedor strain-promoted azide-alkyne ligation methods.

One of the most powerful reagents for introduction of an SO₂F group isethenesulfonyl fluoride (ESF), a strong Michael acceptor as well asDiels-Alder dienophile.

ESF is derived by elimination from 2-chloroethylsulfonyl fluoride, firstdescribed from the sulfonyl chloride in 1932 and reported in large scalewith elimination as a side reaction in 1979. Using the newKFHF-modification (vida supra) in the first stage from ClCH₂CH₂SO₂Cl,ESF can be readily prepared in large quantities (FIG. 8). The relatedlarge-scale preparation of ESF from ethenylsulfonyl chloride (ESCl) waspatented in 1950 by Hedrick (Dow Chemical), but with KF as thenucleophile rather than KFHF, resulting in a relatively low yield (75%).

Several examples of reactions of ESF are shown in FIG. 8. Reactions withactive amines are usually complete within a few minutes at roomtemperature. The participation of amine-containing zwitterions such asamino acids achieves the full level of generality and conveniencerequired of click reactions (FIG. 8, Panel A). One simply stirs a slurryof starting zwitterion in aqueous ethanol, adds the requisite amount ofESF (one molar equivalent for secondary amines like proline, twoequivalents for primary amines), and monitors the stirred suspension forconversion to the new zwitterion. Upon completion, the product isharvested by concentration and filtration. Indeed, for most ESF-amineconjugate additions, purification is rarely required. Details of theimproved preparative procedure for ESF (FIG. 8, top) can be found below.The literature also describes examples of fluorinated derivatives ofESF, which should be similarly useful. ESF is a toxic molecule, sostrict attention to proper procedures for handling this volatilecompound is recommended.

The smallest member of the S(VI) oxyfluorides family, SO₂F₂ was firstdescribed in 1901 by Moissan and subsequently developed by Dow Chemicalin the 1950s as VIKANE pest control agent. At normal temperature andpressure, SO₂F₂ is a colorless, odorless gas, 3.5 times heavier than air(see Table 1). These properties, coupled with its high vapor pressureand ability to saturate air at concentrations lethal to pests, makeSO₂F₂ an effective fumigant, presently used against insects and rodents.The global production of SO₂F₂ since 2000 averages approximately 3million kilograms per year.

TABLE 1 Physical properties of SO₂F₂. CAS number 2699-79-8 molecularweight 102.1 specific gravity 4.18 boiling point −55° C. vapor pressure1611.47 kPa at 20° C. odor odorless appearance colorless gasflammability non-flammable solubility    water = 0.75, (25° C., g/L)1-octanol = 14,   heptane = 22, 1,2-dichloroethane = 25,        MeOH =33,  EtOAc = 59,  acetone = 71

SO₂F₂ is relatively inert in gaseous form and is stable up to 400° C.when dry, but decomposes when heated in air, generating toxic fumes ofHF and SO₂. It is slowly hydrolyzed in water under neutral conditionsand more rapidly under basic conditions to produce fluorosulfate andfluoride ions. SO₂F₂ has relatively small magnetic and quadrupolemoments, does not undergo photolysis in the actinic region of solarradiation, and is inert toward ozone and the active radicals of theatmosphere (Cl., OH.). Again, comparison to sulfuryl chloride isinstructive: SO₂Cl₂ is less thermally stable (decomposes at 100° C. inan open system to chlorine and sulfur dioxide) and easily generateschlorine radicals.

Early published syntheses of fluorosulfates (also called sulfoxylfluorides or sulfurofluoridates; fluorosulfonate is also used althoughthis term should be reserved for compounds containing at least onecarbon-sulfur bond) from phenols used ClSO₂F+SOF₄ or SO₂F₂ at hightemperatures, giving poor results. Chlorosulfates (ROSO₂Cl), unlike theorganic sulfonyl chlorides described above, respond poorly to attemptedsubstitution with KF. Furthermore, chlorosulfates are unattractivestarting materials, as they are prone to self-chlorination and otherradical decomposition processes at low temperatures. The reaction ofSO₂F₂ with preformed sodium and lithium phenolates had previously beenshown to provide better yields of fluorosulfates, but these proceduresdid not catch on. SO₂F₂ therefore represents a curious combination ofton-scale application in the field and poor appreciation in thelaboratory.

Reaction of SO₂F₂ with oxygen nucleophiles in the presence of base givesfluorosulfates (FIG. 10), which have long been known to be quite stabletoward hydrolysis under neutral or acidic conditions. Depending on thenature of the substituent R, the OSO₂F unit can be a good leaving groupor a robust connector. The former reactivity pattern includes theconversion of carboxylic acids and primary alcohols to acyl andaliphatic fluorides, respectively, using SO₂F₂ in the presence of base.Secondary fluorosulfates can be made and isolated when the carbinolcenter is embedded in the molecule between electron-withdrawingsubstituents that make both S_(N)1 and S_(N)2 substitution difficult, asis the case with a C6-fluorosulfate penicillin analogue tested as acovalent inhibitor of porcine pancreatic elastase. In addition, certainperfluorinated aliphatic fluorosulfates can be isolated and were shownto form stable sulfate and sulfamate connections (T. Huang, J. M.Shreeve, Inorg. Chem. 1986, 25, 496-498).

The reaction of SO₂F₂ with alcohols reaches its zenith for aromaticsubstrates, since the derived aryloxyfluorosulfates are very stable.Even more importantly for biological applications, aromatic alcoholsundergo selective modification by SO₂F₂ gas, leaving aliphatic alcohols,aliphatic and aromatic amines, and carboxylates untouched. A widevariety of phenols can be converted to fluorosulfates in quantitativeyields by exposure to gaseous SO₂F₂ and triethylamine (FIG. 11). Forreactions described herein, SO₂F₂ was introduced from a balloon afterthe reaction flask was septum sealed, and the reactions were conductedwith vigorous stirring of the liquid to facilitate gas dissolution inthe condensed phases. The products were isolated by evaporative removalof solvent followed by acidic aqueous extraction to eliminate traces ofbase. Aqueous-organic biphasic conditions were found to suppress, almostcompletely, competitive fluorosulfonation of groups other than phenolsin diversely functionalized molecules such as vancomycin. Thisselectivity for phenolic hydroxyls is remarkable; see FIG. 11, Panel B.Sterically hindered substrates performed best when phenolate anions werepre-formed. Cyclic sulfates are the exclusive products from1,2-catechols under standard reaction conditions, obtained in muchgreater yields than is usually the case with sulfuryl chloride.

Since no reliable methods were previously available for the synthesis offluorosulfates, their chemistry has remained mostly unexplored. Thearyl-sulfate connection (Ar—O—SO₂—) is a vastly underappreciatedlinkage, now formed with sufficient reliability to be applied to a widevariety of targets in biology and materials science. For example,sulfates are phosphate isosteres, and several members of the alkalinephosphatase superfamily can cross-catalyze both phosphoryl and sulfuryltransfer. The reactivity of aryl fluorosulfates towards nucleophiles,including hydroxide, is much diminished compared to the analogoussulfonyl fluorides. Thus, the unassisted reaction of fluorosulfates withsecondary amines requires elevated temperatures in organic solvents. Avery good way to activate these reagents for synthetic chemistry is withtertiary amine catalysts such as DBU as described below. The process canalso be facilitated by vigorous stirring with an immiscible bufferedaqueous phase. The blending of water with miscible cosolvents such asTHE or acetonitrile will also aid the process, but with longer time forcompletion. A benefit of the two-phase process previously noted but notwidely appreciated is that the interfacially-controlled biphasicreactions are usually cleaner than their homogeneous counterparts, evenif rates are similar. Water-tolerant or water-assisted reactions, suchas the addition of nucleophiles to arylfluorosulfates, preferably aretried first in a two-phase format with organic solvent. The productiveinterplay between O₂S—F and F⁻/H⁺ interactions makes this especiallytrue for SuFEx chemistry, as highlighted in FIG. 2.

The synthesis and use of aryl fluorosulfates finds another powerful setof applications when silicon is brought into play. Aryl silyl ethers areexcellent substrates for conversion to fluorosulfates with sulfurylfluoride gas in the presence of catalytic amounts of1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, FIG. 12, reaction A).Trimethylsilyl ethers to give fluorosulfates rapidly (substantiallycomplete within minutes or seconds), whereas the bulkiertert-butyldimethylsilyl group requires several hours for the reaction toreach completion.

In a similar fashion, Lewis bases such as DBU mediate coupling betweensilyl ethers and fluorosulfates, representing the best synthesis ofstable sulfate connections (FIG. 12, reaction B). Arylsulfates aregenerated in high yields with only inert (and sometimes volatile) silylfluorides as byproducts. A wide variety of functional groups can betolerated (FIG. 12), limited only by steric bulk at silicon and thepresence of acidic protons that can quench the basic catalyst.

The conversion of aromatic silyl ethers into fluorosulfates anddiarylsulfates is quite different from the popular use of silylsulfonates (usually triflates) as catalysts in processes such asacetalization, aldol, and allylation reactions. In these and many othercases, the reaction is fundamentally one of electron deficiency, beingaccelerated by the Lewis acidity of silyl sulfonate; the electron-richsilicon component (silyl enol ether or allyl) is nucleophilic enough tocapture an activated intermediate. Silicon-oxygen bonds are swapped, orSi—C is exchanged for Si—O. In the present case, the reaction iselectron-rich, pushed by base catalyst and the ability of sulfuryland/or silicon centers to achieve higher coordination number and becomenucleophilic. The sulfuryl-F bond is strong enough to avoid unwantedside reactions while allowing fluorine to be delivered to silicon as thethermodynamically favored destination.

Another important property of aromatic fluorosulfates is their abilityto participate in transition metal catalyzed coupling reactions (FIG.13). The participation of fluorosulfates as electrophilic components inNegishi and Stille cross-couplings (G. P. Roth, C. E. Fuller, J. Org.Chem. 1991, 56, 3493-3496), as well as palladium catalyzedalkoxycarbonylation reactions ((a) G. P. Roth, J. A. Thomas, TetrahedronLett. 1992, 33, 1959-1962. (b) G. P. Roth, C. Sapino, Tetrahedron Lett.1991, 32, 4073-4076). Competition studies between phenyl fluorosulfateand phenyl triflate showed these groups to have similar coupling rateswith an organotin reagent. Fluorosufate also has utility as aninexpensive triflate alternative (M. A. McGuire, E. Sorenson, F. W.Owings, T. M. Resnick, M. Fox, N. H. Baine, J. Org. Chem. 1994, 59,6683-6686). Fluorosulfate prepared from the corresponding phenol andfluorosulfonic acid anhydride, the most common procedure at the time,engaged in efficient palladium catalyzed methoxycarbonylation on a50-gallon scale.

The replacement of triflate (OTf) with fluorosulfate (OSO₂F) was alsoshown to be practical for enol ethers. Thus, fluorosulfonyl enolatesparticipate in Stille (G. P. Roth, C. Sapino, Tetrahedron Lett. 1991,32, 4073-4076) and Suzuki cross-couplings (L. N. Pridgen, G. K. Huang,Tetrahedron Lett. 1998, 39, 8421-8424), and can also be used asprecursors to allenes (J. Kant, J. A. Roth, C. E. Fuller, D. G. Walker,D. A. Benigni, V. Farina, J. Org. Chem. 1994, 59, 4956-4966) and alkynes(M. Y. Lebedev, E. S. Balenkova, Zh. Org. Khim. 1988, 24, 1156-1161). Wehave found SO₂F₂ to be an effective reagent for the synthesis offluorosulfonyl enol ethers from the related lithium enolate or silylether (FIG. 14).

Primary amines are rapidly fluorosulfonylated by SO₂F₂ gas, but theresulting adducts undergo facile elimination to azasulfene intermediatesby virtue of the acidic nature of the N-sulfamoyl proton. Capture byamine provides symmetrically substituted sulfamides (FIG. 15, Panel A).A few reports are available on the synthesis of monosubstitutedsulfamoyl fluorides by other means. These include Hofmann rearrangementof aryl sulfonamides (FIG. 15, Panel B), halide exchange of parent alkylsulfamoyl chlorides (FIG. 15, Panel C), and ring opening of an aziridineunder fluorinating conditions (FIG. 15, Panel D). All of these processeswere performed under acidic or neutral conditions to avoid theaforementioned elimination of HF from the products.

In contrast, secondary amines react smoothly with SO₂F₂ to giveN-disubstituted sulfamoyl fluorides as remarkably stable compounds,dramatically more robust than analogous chlorides (FIG. 16). Typically,an activating agent such DMAP or DABCO is required, ranging from 0.5equiv. for cyclic amines (exothermic reaction) to a full equivalent foracyclic amines. A variety of solvents can be used, with CH₂Cl₂ or THEproviding the best reaction rates, and the reaction setup is identicalto that described above for the synthesis of fluorosulfates (FIG. 11).The resulting sulfamoyl fluorides are purified by a simple acidic wash.Poor nucleophiles such as disubstituted anilines do not participate inthe reaction with SO₂F₂ under these conditions within a reasonableperiod of time.

We have found N-disubstituted sulfamoyl fluorides to be stable towardhydrolysis under basic conditions at room temperature for more than aweek. Nucleophilic displacement of fluoride in this system requiresheating and some assistance for the departure of fluoride by ahydrogen-bonding solvent, such as water (FIG. 17, top). The reaction islikely to have both S_(N)1 and S_(N)2 character, depending on the natureof the substituents and nucleophile. Furthermore, this type of sulfamoylfluoride is remarkably inert toward a wide range of nucleophiles at roomtemperature in organic solvents, including amines, phosphines, thiols,organolithium and Grignard reagents, hydride, phenoxide, and hydroxide.FIG. 17 shows results from evaluations of the compatibility of theR₂NSO₂F group with a variety of useful synthetic transformations,including those involving strong nucleophiles, reducing agents,oxidants, radicals, and strong acids and bases.

Three key features characterize the SuFEx reactions described herein.First, the SO₂—F bond is unusually strong, so that undesiredsubstitution (such as hydrolysis) is minimized.

This allows precise modifications of complex targets such asbiopolymers. Second, the fluoride radical is inaccessibly energetic, andso radical pathways that complicate the chemistry of other sulfonylhalides do not exist for sulfonyl fluorides. Third, two partners offerversatile ways to make and activate SO₂—F bonds. The proton formsunusually strong hydrogen bonds to fluoride. Even weakly acidicsolvents, additives, and especially interfaces can thereby assist theheterolytic cleavage of the SO₂—F bond. Furthermore, the bifluoride ion(HF₂—) is an excellent source of moderately nucleophilic fluoride forsubstitution reactions. Under non-protic conditions, silicon is useful,as Si and F form the strongest single bond in nature, allowing for therapid formation of SO₂—O bonds from very stable silyl ether precursors.

These factors allow for robust methods for the synthesis of carbon-,oxygen-, and nitrogen-substituted sulfonyl fluorides (sulfonylfluorides, fluorosulfates, and sulfamoyl fluorides), spanning a widerange of stabilities and allowing them to be used in a predictable andpowerful manner in a variety of settings. For synthetic chemists, thefluorosulfate group can function as an inexpensive triflate alternative.With regard to medicinal chemistry, fluorosulfate and sulfamoyl fluoridegroups are useful pharmacophores, and controllable covalent modifiers ofbiomolecules. In all applications, simple, inexpensive, and easilyscalable preparative methods are strongly enabling; we hope that thoseshown here using sulfuryl fluoride gas will spur the development offluorosulfuryl building blocks for many useful purposes. The capacity toignore the irrelevant and respond forcefully to the desired target orcondition makes SO₂F groups particularly useful to probe complexmolecular landscapes such as protein surfaces, or make small-moleculeconnections with absolute reliability.

Hydrolysis and Conversion of ArOSO₂F Groups to ArOH or ArO—SO₃ Groups.

In addition to providing biologically active compounds, the ArOSO₂Fgroup also can be utilized as a simple and selective protecting groupfor any ArOH and ArOSO₃ ⁻ groups or a convenient means for preparingArOSO₃ ⁻ salts. For example, ArOSO₂F compounds can be selectivelyreductively hydrolyzed with aqueous sulfite to afford the correspondingArOH compounds in very high yield. This facile hydrolysis can beachieved by simply stirring the ArOSO₂F compound with an aqueous sulfitesalt such as potassium sulfite, sodium sulfite, and the like (e.g.,about 20 mM to about 2 M sulfite in water). Alternatively, the ArOSO₂Fcompounds can be hydrolyzed to ArOSO₃ ⁻ salts by reaction with anhydrousammonia in methanol in the presence of a carbonate salt such aspotassium carbonate or cesium carbonate (e.g., about 2 molar equivalentsof potassium or cesium carbonate). The reaction to form ArOSO₃ ⁻ israpid and clean, unlike most syntheses of such compounds. For example,the 4-fluorosulfonyloxyphenylacetamide was readily converted to4-hydroxyphenylacetamide and to phenylacetamide-4-sulfate in essentiallyquantitative yields by these procedures.

The following non-limiting examples are provided to further illustratevarious features and aspects of the compositions and methods describedherein.

Example 1 Ex. 1(A). Abbreviations

BEMP=2-tert-butylimino-2-diethylamino-1,3-dimethylperhydro-1,3,2-diazaphosphorine,DBN=1,5-diazabicyclo[4.3.0]non-5-ene,DBU=1,8-diazabicyclo[5.4.0]undec-7-ene, LHMDS=lithiumbis(trimethylsilyl)amide, TCEP=tris(2-carboxyethyl) phosphinehydrochloride, TMS=trimethylsilyl, TBS=tert-butyldimethylsilyl

Ex. 1(B). General Methods

¹H and ¹³C NMR spectra were recorded on Bruker DRX-500, Bruker DRX-600,Bruker AMX-400 instruments and the chemical shifts (6) are expressed inparts per million relative to residual CHCl₃, acetone, acetonitrile orDMSO as internal standards. Proton magnetic resonance (¹H NMR) spectrawere recorded at 600, 500, or 400 MHz. Carbon magnetic resonance (¹³CNMR) spectra were recorded at 150, 125, or 101 MHz. Fluorine magneticresonance (¹⁹F NMR) spectra were recorded at 376 MHz. NMR acquisitionswere performed at 295 K unless otherwise noted. Abbreviations are: s,singlet; d, doublet; t, triplet; q, quartet, p, pentet; br s, broadsinglet. Infrared spectra were recorded as pure undiluted samples usinga THERMONICOLET AVATAR 370 Fourier transform infrared spectrometer witha SMART MIRACLE HATR attachment. Melting points (mp) were determinedusing a THOMAS-HOOVER melting point apparatus and are uncorrected. GC-MSdata were recorded on an AGILENT 7890A GC system with an AGILENT 5975CInert MSD system operating in the electron impact (EI+) mode [Method:T₀=50° C., t=2.25 min; T₁=300° C., ramp=60° C./min, then T₁=300° C., t=4min]. HPLC was performed on an AGILENT 1100 LC/MSD with an Agilent 1100SL mass spectrometer (electrospray ionization, ES) eluting with 0.1%trifluoroacetic acid in H₂O and 0.05% trifluoroacetic acid in CH₃CN.High resolution mass spectrometry was performed on an Agilent ES-TOFinstrument. Precoated MERCK F-254 silica gel plates were used for thinlayer analytical chromatography (TLC) and visualized with short wave UVlight or by potassium permanganate stain. Column chromatography wascarried out employing EMD (Merck) Silica Gel 60 (40-63 m). All startingmaterials were purchased from Alfa Aesar, Aldrich, Acros, AKScientific,Fisher, Lancaster, or TCI chemical companies and used as received.Solvents were purchased from Aldrich, Fisher or Acros chemical companiesand used as received (no extra drying, distillation or special handlingpractices were employed).

Ex. 1(C). Reactions Illustrating the Properties of Sulfonylfluorides Vs.Other Sulfonyl Halides

3-(Fluorosulfonyl)benzoyl chloride (1-1). 3-(Fluorosulfonyl)benzoic acid(2.1 g; 10 mmol) was dissolved in CH₂Cl₂ (20 mL) at room temperature andoxalyl chloride (40 mL of 1M solution in CH₂Cl₂, 2 equiv) was added at0° C., followed by slow addition of DMF (0.2 mL). The reaction wasstirred at room temperature for 6-8 hours. After that time, solvent andvolatiles were removed by rotary evaporation to give3-(fluorosulfonyl)benzoyl chloride as a yellow oil, which was useddirectly in the next step.

3-((3-Ethynylphenyl)carbamoyl)benzene-1-sulfonylfluoride (1-2). Aftercooling to 0° C. on an ice bath, a solution of compound 1-1 in CH₂Cl₂(20 mL) was treated with 3-ethynylaniline (1.2 g, 10 mmol) in CH₂Cl₂ (20mL), followed by slow addition of Et₃N (1.5 mL, 11 mmol, 1.1 equiv). Thereaction mixture was stirred at room temperature overnight, monitored byTLC. After that time, the reaction was washed twice with 1N HCl,extracted with CH₂Cl₂ (100 mL×2). The combined organic layers were driedover Na₂SO₄ and concentrated. The crude product was purified by columnchromatography (3:1 hexane:EtOAc, R_(f) 0.35) to give the desiredproduct as a white solid in 95% yield (2.85 g for 2 steps). mp 155-157°C. ¹H NMR (400 MHz, CDCl₃) δ 8.60 (s, 1H), 8.40 (d, J=8.0 Hz, 1H), 8.24(d, J=8.0 Hz, 1H), 7.88-7.84 (m, 2H), 7.70 (m, 1H), 7.33 (t, J=8.0 Hz,1H), 7.26-7.23 (m, 1H), 3.50 (s, 1H); ¹³C NMR (101 MHz, CDCl₃) δ 165.8,139.7, 138.0, 136.0, 134.7 (d, J=25 Hz), 132.2, 131.6, 130.0, 129.3,128.7, 125.3, 124.3, 122.5, 84.0, 78.9; ¹⁹F NMR (376 MHz, CDCl₃) δ+64.7; ESI-MS (m/z): 304 [MH]⁺.

4-(Bromomethyl)benzene-1-sulfonylfluoride (1-3) was obtained in 93%yield as a white solid (0.9 g). GC-MS analysis revealed contamination ofthe titled compound with 15% of the chloro analogue(4-(chloromethyl)benzene-1-sulfonyl fluoride). This mixture was used inthe next step without purification.

4-(((1R,3S,5S)-3-(4-(((1-Methyl-2-oxo-1,2-dihydroquinolin-4-yl)oxy)methyl)-1H-1,2,3-triazol-1-yl)-8-azabicyclo[3.2.1]octan-8-yl)methyl)benzene-1-sulfonylfluoride(1-4)

Crude 4-(bromomethyl)benzene-1-sulfonyl fluoride (51 mg; 0.2 mmol) andthe 3-triazolyl-8-azabicyclo[3.2.1]octane amine (see Grimster, N. P.;Stump, B.; Fotsing, J. R.; Weide, T.; Talley, T. T.; Yamauchi, J. G.;Nemecz, A.; Kim, C.; Ho, K.-Y.; Sharpless, K. B. J Am Chem Soc 2012,134, 6732-6740.) (73 mg; 0.2 mmol) were mixed in CH₃CN (1 mL) at roomtemperature. Potassium carbonate (30 mg; 1.1 equiv) was added and thereaction mixture was stirred at room temperature overnight, monitored byLC-MS. After completion, the reaction mixture was concentrated by rotaryevaporation and the residue purified by column chromatography (6/1EtOAc/MeOH), giving the desired product as a yellow oil (99 mg, 93%yield). ¹H NMR (400 MHz, CD₃OD) δ 8.28 (s, 1H), 7.97 (d, J=8.0 Hz, 2H),7.82 (d, J=8.0 Hz, 1H), 7.77 (d, J=8.0 Hz, 2H), 7.56 (m, 1H), 7.39 (d,J=8.0 Hz, 1H), 7.16 (t, J=8.0 Hz, 1H), 6.14 (s, 1H), 5.27 (s, 2H), 4.95(m, 1H), 4.90 (s, 1H), 3.81 (s, 2H), 3.38 (m, 2H), 2.34 (t, J=4.0 Hz,2H), 2.22-2.19 (m, 2H), 2.08-2.01 (m, 2H), 1.97 (s, 1H), 1.84 (d, J=12.0Hz, 2H); ¹³C NMR (101 MHz, CD₃OD) δ 165.4, 163.2, 150.0, 143.0, 140.6,132.8, 132.5 (d, J=25 Hz), 131.1, 129.5, 124.5, 124.2, 123.3, 117.3,115.7, 97.6, 63.1, 60.4, 56.1, 54.9, 38.7, 29.7, 27.2; ¹⁹F NMR (376 MHz,CD₃OD) δ +64.6; ESI-MS (m/z): 538 [MH]⁺.

Ex. 1(D). Demonstration of Hydrolytic Stability of SulfonimidoylFluorides

Sulfonimidoyl fluoride (1 mmol) was dissolved in acetonitrile (0.1 M),diluted with equal amount of buffer to initiate the hydrolysis process,and stirred at room temperature for several hours. Aliquots were takenat time periods and the composition of the mixture analyzed by massspectrometry. FIG. 19 shows the loss of starting fluoride as a functionof time.

Ex. 1(E). Sulfonylfluorides Made with Potassium Bifluoride (See FIG. 19)Representative Procedure for the Synthesis of Sulfonylfluorides UsingSaturated Aqueous KHF₂ and CH₃CN. 4-Bromobenzene-1-Sulfonylfluoride(5-1)

KHF₂ (71 g; 2.3 equiv) was dissolved in H₂O (200 mL) to make a saturatedsolution (endothermic reaction), which was treated with a solution of4-bromobenzene-1-sulfonyl chloride (100 g, 1 equiv) in acetonitrile (400mL). The reaction mixture was stirred at room temperature for 3 h,monitored by GC-MS. After that time, the reaction mixture wastransferred to a separatory funnel and the organic layer was collected.The aqueous phase was extracted with EtOAc (300 mL), and the combinedorganic extracts were washed with 10% aqueous NaCl (2×), saturatedsodium chloride (1×), dried over sodium sulfate, and concentrated byrotary evaporation to give the desired product as a white solid (91.5 g,98% yield). mp 63-64° C. ¹H NMR: (400 MHz, CDCl₃) δ 7.88 (dd, J=9.2 Hz,2 Hz, 2H), 7.78 (dd, J=9.2 Hz, 2 Hz, 2H); ¹³C NMR: (101 MHz, CDCl₃) δ133.1, 131.9 (d, J=25.5 Hz), 131.3, 129.8; ¹⁹F NMR (376 MHz, CDCl₃) δ+65.9; GC-MS: 4.68 min; EI (m/z): 240/238 [M]⁺.

None of the sulfonyl fluoride compounds prepared by this procedurerequired extra purification.

2-Bromobenzene-1-sulfonylfluoride (5-2): light yellow crystals (1.1 g,97% yield). mp 48-49° C. ¹H NMR (400 MHz, CDCl₃) δ 8.12 (dd, J=7.2 Hz,2.4 Hz, 1H), 7.84 (dd, J=7.2 Hz, 1.6 Hz, 1H), 7.56 (m, 2H); ¹³C NMR (101MHz, CDCl₃) δ 136.1, 135.9, 133.8 (d, J=23.9 Hz), 132.0, 127.9, 121.0;¹⁹F NMR (376 MHz, CDCl₃) δ +57.4; GC-MS (t_(R)): 34.8 min; EI-MS (m/z):240/238 [M]⁺.

2-Methylbenzene-1-sulfonylfluoride (5-3): colorless oil (1.54 g, 90%yield). ¹H NMR (400 MHz, CDCl₃) δ 8.01 (d, J=8 Hz, 1H), 7.64-7.60 (m,1H), 7.43-7.38 (m, 2H), 2.68 (s, 3H); ¹³C NMR (101 MHz, CDCl₃) δ 139.0,135.4, 132.9, 132.3 (d, J=22.2 Hz), 130.0, 126.7, 20.2; ¹⁹F NMR (376MHz, CDCl₃) δ +59.8; GC-MS (t_(R)): 4.1 min; EI-MS (m/z): 174 [M]⁺.

4-(Fluorosulfonyl)benzoic acid (5-4): white solid (6.9 g, 90% yield)prepared by the above procedure using THE instead of acetonitrile. mp273-274° C. ¹H NMR (400 MHz, CD₃OD-d₄) δ 8.21-8.18 (m, 2H), 8.07-8.03(m, 2H); ¹³C NMR (101 MHz, CD₃OD-d₄) δ 167.3, 138.9, 137.6 (d, J=25.3Hz), 132.0, 129.7; ¹⁹F NMR (376 MHz, CD₃OD-d₄) δ +64.2; EI-MS (m/z): 205[M]⁺.

3-(Fluorosulfonyl)benzoic acid (5-5): white solid (19.5 g, 96% yield)prepared by the above procedure using THE instead of acetonitrile. mp152-154° C. ¹H NMR (400 MHz, CD₃OD-d₄) δ 8.45 (s, 1H), 8.33 (d, J=8 Hz,1H), 8.12 (d, J=8 Hz, 1H), 7.71 (t, J=8 Hz, 1H); ¹³C NMR (101 MHz,CD₃OD-d₄) δ 166.9, 137.7, 134.7 (d, J=25.3 Hz), 134.2, 133.2, 131.7,130.3; ¹⁹F NMR (376 MHz, CD₃OD-d₄) δ +64.5; EI-MS (m/z): 205 [M]⁺.

4-Nitrobenzene-1-sulfonylfluoride (5-6): white solid (22.6 g, 97%yield). mp 75-76° C. ¹H NMR (400 MHz, CDCl₃) δ 8.49 (d, J=8 Hz, 2H),8.25 (d, J=8 Hz, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 151.9, 138.4 (d, J=27.3Hz), 130.1, 125.0; ¹⁹F NMR (376 MHz, CDCl₃) δ +66.0; GC-MS (t_(R)): 4.9min; EI-MS (m/z): 205 [M]⁺.

3-Nitrobenzene-1-sulfonylfluoride (5-7): white solid (22.3 g, 97%yield). mp 42-43° C. ¹H NMR (400 MHz, CDCl₃) δ 8.86 (t, J=2.4 Hz, 1H),8.64 (d, J=8 Hz, 1H), 8.35 (d, J=8 Hz, 1H), 7.93 (t, J=8 Hz, 1H); ¹³CNMR (101 MHz, CDCl₃) δ 148.4, 134.9 (d, J=28.3 Hz), 133.8, 131.4, 130.0,123.9; ¹⁹F NMR (376 MHz, CDCl₃) δ +65.9; GC-MS (t_(R)): 4.85 min; EI-MS(m/z): 205 [M]⁺.

2-Nitrobenzene-1-sulfonylfluoride (5-8): yellow solid (2.1 g, 98%yield). mp 55-57° C. ¹H NMR (400 MHz, CDCl₃) δ 8.23 (dd, J=8 Hz, 1.6 Hz,1H), 8.04 (d, J=8 Hz, 1H), 7.97 (dt, J=8 Hz, 1.6 Hz, 1H), 7.88 (m, 1H);¹³C NMR (101 MHz, CDCl₃) δ 136.7, 133.4, 131.7, 126.8 (d, J=28.6 Hz),125.8; ¹⁹F NMR (376 MHz, CDCl₃) δ +64.5; GC-MS (t_(R)): 5.2 min; EI-MS(m/z): 205 [M]⁺.

Benzene-1,2-disulfonyl difluoride (5-9): white solid (0.87 g, 100%yield). mp 126-128° C. ¹H NMR (400 MHz, CDCl₃) δ 8.44 (dd, J=5.6 Hz, 3.2Hz, 2H), 8.07 (dd, J=6 Hz, 3.2 Hz, 2H); ¹³C NMR (101 MHz, CDCl₃) δ136.1, 133.5, 132.2 (d, J=28.7 Hz); ¹⁹F NMR (376 MHz, CDCl₃) δ +65.3;GC-MS (t_(R)): 5.15 min; EI-MS (m/z): 242 [M]⁺.

4-Iodobenzene-1-sulfonylfluoride (5-10): white solid (1.4 g, 92% yield).mp 84-85° C. ¹H NMR (400 MHz, CDCl₃) δ 8.02 (d, J=8.8 Hz, 2H), 7.71 (d,J=8.8 Hz, 2H), 7.84 (d, J=8.8 Hz, 2H); ¹³C NMR (101 MHz, CDCl₃) δ139.03, 132.5 (d, J=25.5 Hz), 129.4, 104; ¹⁹F NMR (376 MHz, CDCl₃) δ+65.6; GC-MS (t_(R)): 5.0 min; EI-MS (m/z): 286 [M]⁺.

2-Cyanobenzene-1-sulfonylfluoride (5-11): white solid (0.97 g, 97%yield). mp 87-89° C. ¹H NMR (400 MHz, CDCl₃) δ 8.23-8.21 (m, 1H),8.03-8.01 (m, 1H), 7.98-7.92 (m, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 135.8,135.7, 134.7 (d, J=26.5 Hz), 133.6, 130.8, 114.1, 111.5; ¹⁹F NMR (376MHz, CDCl₃) δ +64.0; GC-MS (t_(R)): 5.0 min; EI-MS (m/z): 185 [M]⁺.

4-(Trifluoromethyl)benzene-1-sulfonylfluoride (5-12): white crystallinesolid (1.0 g, 98% yield). mp 68-69° C. ¹H NMR (400 MHz, CDCl₃) δ 8.17(d, J=8.4 Hz, 2H), 7.92 (d, J 8.4 Hz, 2H); ¹³C NMR (101 MHz, CDCl₃) δ137.0 (q, J=33.5 Hz), 136.4 (d, J=26.7 Hz), 129.1, 126.9 (q, J=3.7 Hz),122.7 (q, J=271.7 Hz); ¹⁹F NMR (376 MHz, CDCl₃) δ +65.4, −64.0; GC-MS(t_(R)): 3.7 min; EI-MS (m/z): 228 [M]⁺.

2,4-Dimethoxybenzene-1-sulfonyl fluoride (5-13): white solid (2 g, 94%yield). mp 108-109° C. ¹H NMR (400 MHz, CDCl₃) δ 7.41 (d, J=3.2 Hz, 1H),7.24 (dd, J=9.2 Hz, 3.2 Hz, 1H), 7.04 (d, J=9.2 Hz, 1H), 3.95 (s, 3H),3.81 (s, 3H); ¹³C NMR (101 MHz, CDCl₃) δ 152.9, 152.2, 123.6, 121.3 (d,J=23.3 Hz), 114.9, 114.3, 56.9, 56.1; ¹⁹F NMR (376 MHz, CDCl₃) δ +58.6;GC-MS (t_(R)): 6.0 min; EI-MS (m/z): 236 [M]⁺.

(E)-Methyl 3-(4-(fluorosulfonyl)phenyl)acrylate (5-14): white solid (0.3g, 93% yield). mp 138-139° C. ¹H NMR (400 MHz, CDCl₃) δ 8.01 (d, J=8.4Hz, 2H), 7.74 (d, J 8.4 Hz, 2H), 7.69 (d, J=16 Hz, 1H), 6.58 (d, J=16Hz, 1H), 3.83 (s, 3H); ¹³C NMR (101 MHz, CDCl₃) δ 166.2, 141.6, 141.4,133.6 (d, J=25.1 Hz), 129.0, 128.7, 122.8, 52.8; ¹⁹F NMR (376 MHz,CDCl₃) δ +65.6; GC-MS (t_(R)): 5.9 min; EI-MS (m/z): 244 [M]⁺.

2,4,6-Triisopropylbenzene-1-sulfonylfluoride (5-15): white solid (0.6 g,100% yield). mp 67-68° C. ¹H NMR (400 MHz, CDCl₃) δ 7.25 (s, 2H), 3.96(dq, J=6.8 Hz, 2.4 Hz, 2H), 2.95 (q, J=6.8 Hz, 1H), 1.30 (d, J=6.8 Hz,12H), 1.27 (d, J=6.8 Hz, 6H); ¹³C NMR (101 MHz, CDCl₃) δ 155.3, 150.7,128.0 (d, J=18.4 Hz), 124.0, 34.4, 30.1, 24.5, 23.4; ¹⁹F NMR (376 MHz,CDCl₃) δ +72.2; GC-MS (t_(R)): 5.7 min; EI-MS (m/z): 286 [M]⁺.

Phenylmethanesulfonyl fluoride (PMSF) (5-16): white solid (1.7 g, 98%yield). mp 93-94° C. ¹H NMR (400 MHz, CDCl₃) δ 7.46-7.44 (m, 5H), 4.59(d, J=4 Hz, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 130.8, 130.0, 129.4, 125.6,56.9 (d, J=17.2 Hz); ¹⁹F NMR (376 MHz, CDCl₃) δ +50.5; GC-MS (t_(R)):4.5 min; EI-MS (m/z): 174 [M]⁺.

3-Chloropropane-1-sulfonylfluoride (5-17): light yellow oil (3.27 g, 98%yield) prepared by the above procedure but omitting acetonitrile (thereaction was performed in suspension on water). ¹H NMR (400 MHz, CDCl₃)δ 3.70 (t, J=6.4 Hz, 2H), 3.58 (dt, J=4.8 Hz, 7.6 Hz, 2H), 2.40 (m, 2H);¹³C NMR (101 MHz, CDCl₃) δ 48.0 (d, J=17.7 Hz), 41.6, 26.4; ¹⁹F NMR (376MHz, CDCl₃) δ +53.9; GC-MS (t_(R)): 3.7 min; EI-MS (m/z): 160 [M]⁺.

((1R,4R)-7,7-Dimethyl-2-oxobicyclo[2.2.2]heptan-1-yl)methanesulfonylfluoride (5-18): colorless crystals (1.2 g, 94% yield). mp 78-80° C. ¹HNMR (400 MHz, CDCl₃) δ 3.82 (dd, J=15.2 Hz, 2.8 Hz, 1H), 3.29 (dd,J=15.2 Hz, 2.8 Hz, 1H), 2.45-2.29 (m, 2H), 2.16 (t, J=4.4 Hz, 1H), 2.07(m, 1H), 1.98 (d, J=18.8 Hz, 1H), 1.73 (m, 1H), 1.48 (m, 1H), 1.11 (s,3H), 0.90 (s, 3H); ¹³C NMR (101 MHz, CDCl₃) δ 213.0, 57.7, 48.3, 48.1(d, J=19.4 Hz), 42.8, 42.2, 26.7, 25.1, 19.5; ¹⁹F NMR (376 MHz, CDCl₃) δ+63.7; GC-MS (t_(R)): 5.34 min; EI-MS (m/z): 234 [M]⁺.

Ex. 1(F). Sulfonylfluorides Made from Sulfonic Acids

Representative procedure for the synthesis of3-azidopropane-1-sulfonylfluoride (6-1). Sodium3-azidopropane-1-sulfonate was obtained as a white solid (16.25 g, 87%yield) according to the procedure in Kong, X., et al. US 2008/0146642A1.

Sodium 3-azidopropane-1-sulfonate (9.35 g, 50 mmol) was suspended intrifluoromethylbenzene (20 mL, 2.5M). Oxalyl chloride (4.3 mL; 50 mmol)was added to the reaction mixture at 0° C., followed by 5 drops of DMF.The resulting white suspension was stirred at room temperature under adry atmosphere (nitrogen or a drying tube packed with desiccant). Thereaction mixture was then added to a cold saturated aqueous solution ofKHF₂ (approx. 4.5 M, 2.5 equiv) in a plastic bottle and the biphasicsuspension was stirred at room temperature. GC-MS analysis after fivehours showed full conversion, and the mixture was filtered into aseparatory funnel using extra organic solvent to wash the solidmaterial. The organic phase was separated, washed with brine, and driedover anhydrous sodium sulfate. The separated aqueous phase wasback-extracted with trifluoromethylbenzene, and the organic phase wasdried and combined with the primary organic solution. Since3-azidopropane-1-sulfonyl fluoride is a volatile compound, the solutionwas stored and used as a stock solution. The concentration of sulfonylfluoride was established as 16 wt-% by quantitative ¹H NMR against thesolvent, consistent with a quantitative yield. The analogous procedurewas performed using 0.5M CH₂Cl₂ in MeCN as the solvent, a 1Mconcentration of 3-azidopropane-1-sulfonate (25 mmol scale), and 1.5equiv. of oxalyl chloride gave the sulfonyl fluoride 6-1 in 87% yield(23 wt-% stock solution) measured by ¹H NMR integration. ¹H NMR (400MHz, CDCl₃) δ 3.54 (t, J=6.3 Hz, 2H), 3.51-3.45 (m, 2H), 2.19 (dq,J=8.8, 6.4 Hz, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 48.9, 48.1 (d, J=17.6Hz), 23.5; ¹⁹F NMR (376 MHz, CDCl₃) δ 53.7; R_(f) (9:1 hexane:EtOAc):0.22; GC (t_(R)): 3.8 min; EI-MS (m/z): 139 [M-28]⁺.

3-Azidobutane-1-sulfonylfluoride (6-2). Sodium 3-azidobutane-1-sulfonatewas obtained as a white solid (21.6 g, 90% yield) according to theprocedure in Kong, X., et al. US 2008/0146642 A1. Here, the above methodin trifluoromethylbenzene gave a 74% yield of 6-2, whereas the reactionin 0.5M CH₂Cl₂ in MeCN provided the desired product in 92% yield (30wt-% stock solution). ¹H NMR (400 MHz, CDCl₃) δ 3.45-3.40 (m, 2H), 3.38(t, J=6.3 Hz, 2H), 2.17-2.04 (m, 2H), 1.84-1.72 (m, 2H); ¹³C NMR (101MHz, CDCl₃) δ 50.5, 50.3 (d, J=17.1 Hz), 27.1, 21.1; ¹⁹F NMR (376 MHz,CDCl₃) δ 53.2; R_(f) (9:1 hexane:EtOAc): 0.28; GC (t_(R)): 5.0 min;EI-MS (m/z): 153 [M-28]⁺.

Pent-4-yne-1-sulfonylfluoride (6-3). Pent-4-yne-1-chloride (0.2 mol) andNa₂SO₃ (0.2 mol) were heated in water (200 mL) at 95° C. for 16 h. Thesolution was concentrated and dried under vacuum to provide a mixture ofsodium pent-4-yne-1-sulfonate and NaCl. This material was used in theabove procedure without further purification. The reaction intrifluoromethylbenzene (25 mmol scale) gave a 50% yield (three stepsfrom pent-4-yne-1-chloride) of the sulfonyl chloride (as a 5.3 wt-%stock solution), whereas the reaction in 0.5M CH₂Cl₂ in MeCN providedthe desired product, 6-3, in 60% yield (three steps, 27 wt-% solution).¹H NMR (400 MHz, CDCl₃) δ 3.63-3.56 (m, 2H), 2.48 (td, J=6.7, 2.6 Hz,2H), 2.28-2.17 (m, 3H); ¹³C NMR (101 MHz, CDCl₃) δ 80.8, 70.8, 49.5 (d,J=17.6 Hz), 22.6, 16.8; ¹⁹F NMR (376 MHz, CDCl₃) δ 53.4; R_(f) (9:1hexane:EtOAc): 0.30.

4-(3-Bromopropyl)benzene-1-sulfonylfluoride (6-4). A solution of3-bromopropyl)benzene (1.1 g) in CHCl₃ (10 mL) was cooled to 0° C. on anice bath and was treated with chlorosulfuric acid (2.2 mL, 6 equiv)slowly by syringe with continued cooling. After 30 min, the ice bath wasremoved and the reaction mixture was stirred at room temperatureovernight. The mixture was poured into an ice-water mixture (100 g) andthe crude sulfonyl chloride was extracted with EtOAc (2×30 mL). Thecombined organic extracts were washed with brine and concentrated. Theresulting crude sulfonyl chloride was dissolved in acetonitrile (10 mL)and treated with saturated aqueous KHF₂ (5 mL). The reaction mixture wasstirred at room temperature overnight, monitoring by GC-MS. Uponcompletion, the sulfonyl fluoride was extracted with EtOAc (3×20 mL),and the combined organic phases were dried over anhydrous Na₂SO₄ andconcentrated to give 6-4 as a yellow oil (1.4 g, 90% yield for twosteps). ¹H NMR (400 MHz, CDCl₃) δ 7.92 (d, J=8.0 Hz, 2H), 7.47 (d, J=8.0Hz, 2H), 3.39 (t, J=8.0 Hz, 2H), 2.92 (t, J=8.0 Hz, 2H), 2.20 (m, 2H);¹³C NMR (101 MHz, CDCl₃) δ 149.7, 130.8 (d, J=25 Hz), 129.9, 128.7,34.0, 33.4, 32.4; ¹⁹F NMR (376 MHz, CD₃OD) δ +65.8; GC-MS (t_(R)): 5.8min; EI-MS (m/z): 280 [M]⁺.

4-Hydroxybenzene-1-sulfonylfluoride (6-5). A 50 mL round-bottom flaskwas charged with 4-hydroxybenzenesulfonate (2.56 g). Thionyl chloride(10 mL) was added and the mixture was heated to reflux under a nitrogenatmosphere for 6 hours. The bulk of the excess thionyl chloride wasremoved by distillation, and the last traces were removed by addition oftoluene (10 mL) and rotary evaporation. The resulting crude sulfonylchloride (a yellow oil) was dissolved in CH₃CN (20 mL) and treated withsaturated aqueous KHF₂ (5 mL). The reaction mixture was stirred at roomtemperature overnight and monitored by GC-MS. Upon completion, thesulfonyl fluoride was extracted with EtOAc (3×20 mL), dried overanhydrous Na₂SO₄, and concentrated. The desired product was obtained ascolorless crystals (1.84 g, 80% yield for two steps). ¹H NMR (400 MHz,CDCl₃) δ 7.86 (d, J=12.0 Hz, 2H), 7.04 (m, 2H); ¹³C NMR (101 MHz, CDCl₃)δ 162.7, 131.2, 123.2 (d, J=23 Hz), 116.7; ¹⁹F NMR (376 MHz, CDCl₃) δ+66.8; ESI-MS (m/z): 177 [MH]⁺.

Ex. 1(G). Sulfonimidoyl Fluorides Prepared from Corresponding Chlorides

General procedure: The starting sulfonimidoyl chloride was dissolved inacetonitrile (0.33 M) and an equal volume of saturated aqueous KHF₂ wasadded with stirring. The reaction mixture was stirred at roomtemperature for several hours. Upon completion (monitored by LC-MS orGC), the acetonitrile layer was separated, dried over Na₂SO₄ andconcentrated. In cases in which crude sulfonimidoyl chlorides were used,pure samples of the product fluoride were obtained by columnchromatography.

N-(Methylsulfonyl)benzenesulfonimidoyl fluoride (7-1) was isolated as awhite solid in 95% yield (0.9 g). mp 77-78° C.; ¹H NMR (500 MHz, CD₃CN)δ 8.15 (d, J=8.0 Hz, 2H), 7.93 (t, J=7.5 Hz, 1H), 7.76 (t, J=8.0 Hz,2H), 3.28 (s, 3H); ¹³C NMR (126 MHz, CD₃CN) δ 137.9, 133.4 (d, J=19.9Hz), 131.3, 128.9, 45.0; ¹⁹F NMR (376 MHz, CDCl₃) δ 71.9; R_(f) (7:3hexane:EtOAc): 0.31; ESI-MS (m/z): 238 [MH]⁺.

N-Tosylbenzenesulfonimidoyl fluoride (7-2). Starting from the crudesulfonimidoyl chloride, the crude fluoride product was purified by shortcolumn chromatography (9:1 hexane:EtOAc to 1/1) to give a white solid(3.8 g, 71% yield). mp 60-61° C.; ¹H NMR (400 MHz, CDCl₃) δ 8.04 (d,J=8.5 Hz, 2H), 7.94 (d, J=8.2 Hz, 2H), 7.79 (t, J=7.5 Hz, 1H), 7.62 (t,J=7.9 Hz, 2H), 7.33 (d, J=8.0 Hz, 2H), 2.43 (s, 3H); ¹³C NMR (1 MHz,CDCl₃) δ 144.5, 139.0, 136.3, 133.3 (d, J=20.2 Hz), 129.9, 129.8, 128.2,127.2, 21.8; ¹⁹F NMR (376 MHz, CDCl₃) δ 73.5; R_(f) (6:4 hexane:EtOAc):0.46; ESI-MS (m/z): 294 [MH]⁺.

N-((3-Azidopropyl)sulfonyl)benzenesulfonimidoyl fluoride (7-3). Startingfrom the crude sulfonimidoyl chloride, the crude fluoride product waspurified by short column chromatography (1:1 CH₂Cl₂:hexane) to give acolorless oil (1.32 g, 79% yield). ¹H NMR (500 MHz, CD₃CN) δ 8.13 (d,J=6.9 Hz, 2H), 7.96-7.88 (m, 1H), 7.78-7.70 (m, 2H), 3.53-3.46 (m, 2H),3.46-3.39 (m, 2H), 2.17-2.07 (m, 2H); ¹³C NMR (126 MHz, CD₃CN) δ 137.9,133.3 (d, J=19.6 Hz), 131.2, 128.9, 54.4, 50.0, 24.3; ¹⁹F NMR (376 MHz,CDCl₃) δ 72.9; R_(f) (6:4 hexane:EtOAc): 0.49; ESI-MS (m/z): 329 [MNa]⁺.

N-Methylcarbonatebenzenesulfonimidoyl fluoride (7-4). The product wasisolated as yellow oil in 82% yield (0.9 g). ¹H NMR (400 MHz, CDCl₃) δ8.09-8.05 (m, 2H), 7.78-7.72 (m, 1H), 7.63-7.57 (m, 2H), 3.78 (s, 3H);¹³C NMR (101 MHz, CDCl₃) δ 154.3, 135.8, 133.3 (d, J=20.6 Hz), 129.7,128.1, 54.2; ¹⁹F NMR (376 MHz, CDCl₃) δ 68.5; R_(f) (8:2 hexane:EtOAc):0.40; ESI-MS (m/z): 218 [MH]⁺.

General procedure. Sulfonimidoyl fluorides with alkyl or arylsubstituents on nitrogen were prepared according to the sequencepreviously described by Kowalczyk, R.; Edmunds, A. J. F.; Hall, R. G.;Bolm, C. Org. Lett. 2011, 13, 768-771. The starting sulfonimidoyl orsulfamoyl chlorides were dissolved in acetonitrile (0.1 M) and treatedwith AgF (1.2-1.5 equiv) in a foil-wrapped flask for 1 hour. Uponcompletion, the reaction was quenched with 1M HCl (0.2 M) and stirred atroom temperature for 30-60 min, then filtered through CELITE, washedwith CH₂Cl₂, concentrated, and purified by short silica gel columnchromatography.

N-(3-Ethynylphenyl)benzenesulfonimidoyl fluoride (7-5) was isolated as abrown oil (0.5 g, 80% yield). ¹H NMR (600 MHz, CD₃CN) δ 8.22-8.19 (m,2H), 7.85 (t, J=7.5 Hz, 1H), 7.72 (t, J=7.9 Hz, 2H), 7.38-7.34 (m, 2H),7.31-7.30 (m, 1H), 7.30-7.28 (m, 1H), 3.43 (s, 1H); ¹³C NMR (151 MHz,CD₃CN) δ 140.6 (d, J=5.4 Hz), 136.4, 136.3, 135.6 (d, J=24.8 Hz), 130.8,129.0, 128.6, 127.9 (d, J=5.0 Hz), 125.5 (d, J=4.6 Hz), 124.1, 83.5,79.4; ¹⁹F NMR (376 MHz, CD₃CN) δ 80.3; R_(f) (6:4 hexane:EtOAc): 0.47;ESI-MS (m/z): 260 [MH]⁺.

N-(1-Ethynylcyclohexyl)-4-nitrobenzene-1-sulfonimidoyl fluoride (7-6).The starting sulfonimidoyl chloride was prepared starting from thesulfenyl amide, followed by oxidation with mCPBA. Using the abovegeneral procedure, 7-6 was isolated as a yellow oil 1.0 g, 81% yield).¹H NMR (400 MHz, CDCl₃) δ 8.37 (d, J=8.8 Hz, 2H), 8.23 (d, J=8.8 Hz,2H), 2.55 (s, 1H), 2.15-2.04 (m, 2H), 1.92-1.83 (m, 2H), 1.72-1.61 (m,4H), 1.59-1.50 (m, 1H), 1.37-1.26 (m, 1H); ¹³C NMR (101 MHz, CDCl₃) δ150.8, 142.5 (d, J=30.2 Hz), 142.4, 129.1, 124.3, 86.1 (d, J=5.8 Hz),72.3, 56.4 (d, J=3.2 Hz), 41.0 (d, J=2.2 Hz), 40.8 (d, J=4.0 Hz), 25.1,22.9 (d, J=3.8 Hz); ¹⁹F NMR (376 MHz, CDCl₃) δ 90.5; R_(f) (7:3hexane:EtOAc): 0.68; ESI-MS (m/z): 311 [MH]⁺.

Ex. 1(H). Preparation of 2-Chloroethanesulfonyl Fluoride and ESF(Adapted from Hyatt et al. JOC, 1979, 44, 3847-3858)

All manipulations with 2-chloroethane-1-sulfonyl chloride,2-chloroethane-1-sulfonyl fluoride, and ESF must be performed in awell-ventilated fume hood. As powerful alkylating agents, care must betaken to avoid inhalation or skin contact.

2-Chloroethanesulfonyl fluoride. A 1 L round-bottom flask was chargedwith a magnetic stirring bar, KHF₂ (187 g, 2.4 mol) and water (0.5 L).The reaction mixture was stirred at room temperature for 20 min untilcomplete dissolution of KHF₂ was achieved (this is an endothermicprocess, reaching an internal temperature of 5° C.).2-Chloroethane-1-sulfonyl chloride (180.7 g, 1.05 mol) was poured intothe cold saturated potassium bifluoride solution, and the reactionmixture was vigorously stirred at room temperature for 3 hours. Thereaction was monitored by GC-MS (using a low injection and columntemperature) or by NMR. Upon completion, the organic layer of the neatsulfonyl fluoride was separated and washed with brine, givingapproximately 150 g of 2-chloroethane-1-sulfonyl fluoride as a lightyellow oil. The rest of the product was extracted from the aqueous KHF₂layer with CH₂Cl₂ (100 mL). This organic extract was washed with brine,concentrated, and combined with the first batch of product. ¹H NMR (400MHz, CDCl₃) δ 3.94-3.90 (m, 2H), 3.83-3.78 (m, 2H); ¹³C NMR (101 MHz,CDCl₃) δ 52.5 (d, J=7.2 Hz), 35.0; ¹⁹F NMR (376 MHz, CDCl₃) δ +57.2;GC-MS (t_(R)): 2.3 min; EI-MS (m/z): 146 [M]⁺.

The recovered bifluoride solution (now containing a mixture of KH₂F₃ andKCl) was recharged by adding 1 equiv KF solution. The above reaction wasrepeated using this solution at 0.5 mol scale, providing2-chloroethanesulfonyl fluoride in 100% yield. Two morere-charge/reaction cycles at the same scale each gave 98% yields.

Ethenesulfonyl fluoride (ESF). A 1 L 2-necked round-bottom flaskequipped with stirring bar and internal thermometer was placed in anice-water bath. The reaction flask was charged with ice-cold water (0.4L) and 2-chloroethane-1-sulfonyl fluoride (232.5 g; 1.59 mol; 1 equiv).MgO (35.3 g; 0.875 mol; 0.55 equiv) was added portion-wise to thevigorously stirred reaction mixture, keeping the reaction temperaturebelow 35° C., with optimal temperature being about 25° C. (Highertemperatures result in decreased yields, lower reaction temperaturesslow down the reaction too much). After final portion of MgO was added,the reaction mixture was stirred for an additional 2-3 hours. Uponcompletion (monitored by GC, NMR), the bottom layer of neat ESF wasremoved in a separatory funnel. The product was dried by stirring withMgSO₄ for 20 min and filtered giving 125.7 g of neat ESF (72% yield),which was stored in a plastic bottle. The rest of the product (about 2.3g) was extracted from the aqueous layer with dichloromethane (150 mL)and dried over MgSO₄. The resulting solution can be either used as astock solution, or can be concentrated under low-pressure vacuum to giveneat ESF.

Ex. 1(I). ESF Decorations of Nitrogen, Oxygen, and Carbon Nucleophiles

Section A: (S)-1-(2-(Fluorosulfonyl)ethyl)pyrrolidine-2-carboxylic acid(10-1). Proline (5 g; 43 mmol) was suspended in 95:5 EtOH:H₂O (100 mL)and treated with ESF (4 mL, 44 mmol). The reaction mixture was stirredat room temperature for several hours. Upon completion, the yellowsolution was concentrated and dried under vacuum to give the product aswhite solid (9.45 g, 97% yield). ¹H NMR (600 MHz, DMSO-d₆) δ 4.18-4.03(m, ₂H), 3.35-3.28 (m, 1H), 3.25 (dt, J=13.6, 7.0 Hz, 1H), 3.04-2.94 (m,2H), 2.55-2.51 (m, 1H), 2.07-1.97 (m, 1H), 1.87-1.79 (m, 1H), 1.77-1.67(m, 2H); ¹³C NMR (151 MHz, CDCl₃) δ 174.4, 64.2, 51.9, 48.9 (d, J=11.5Hz), 28.7, 23.0; ¹⁹F NMR (376 MHz, CDCl₃) δ +57.6; ESI-MS (m/z): 226[MH]⁺.

(7R)-7-((R)-2-(Bis(2-(fluorosulfonyl)ethyl)ammonio)-2-(4-hydroxyphenyl)acetamido)-3-methyl-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate(10-2). Cefadroxil (363 mg; 1 mmol) was suspended in absolute EtOH (2mL) and treated with ESF (0.2 mL; 2.2 mmol). The reaction mixture wasstirred at 50° C. overnight. The resulting yellow solution wasconcentrated by rotary evaporation and the residue purified by columnchromatography (90:10:3 to 90:10:6 EtOAc:EtOH:H₂O) to give the desiredproduct as a white solid (0.5 g, 86% yield). mp 225-230° C. (dec.); ¹HNMR (400 MHz, DMSO-d₆) δ 9.56 (bs, 1H), 9.00 (d, J=8 Hz, 1H), 7.13 (d,J=8 Hz, 2H), 6.70 (d, J=8 Hz, 2H), 5.59 (t, J=8 Hz, 1H), 5.00 (d, J=4Hz, 1H), 4.70 (s, 1H), 4.13-4.06 (m, 2H), 3.90-3.82 (m, 2H), 3.46 (d,J=16 Hz, 1H), 3.23-3.18 (m, 3H), 3.06-2.99 (m, 2H), 1.94 (s, 3H); ¹³CNMR (101 MHz, DMSO-d₆) δ 172.0, 163.7, 157.7, 130.3, 126.5, 115.6, 66.5,58.8, 57.4, 49.3 (d, J=10 Hz), 45.1, 29.3, 19.8; ¹⁹F NMR (376 MHz,DMSO-d₆) δ +57.9; R_(f) (EtOAc/EtOH/H₂O—90/10/6): 0.41; ESI-MS (m/z):606 [MNa]⁺.

Section B: General procedure for the reaction of primary and secondaryamines with ESF (adapted from Krutak, J. J.; Burpitt, R. D.; Moore, W.H.; Hyatt, J. A. J. Org. Chem. 1979, 44, 3847-3858). The starting amine(1 equiv) was dissolved in organic solvent (usually CH₂Cl₂ or THF,0.1-0.5 M in substrate) and treated with ESF (1-2.5 equiv). The reactionmixture was stirred at room temperature for several minutes to severalhours, monitoring conversion by LC-MS. Upon completion, the solvent andexcess of ESF were removed by rotary evaporation and dried under vacuum,usually providing clean product. When purification by columnchromatography is mentioned, it was done to remove trace impurities.

2,2′-((2-(1H-Indol-3-yl)ethyl)azanediyl)diethanesulfonyl fluoride(10-3). Reaction in CH₂Cl₂; the product was further purified by flashcolumn chromatography (9:1 to 6:4 hexane:EtOAc) to obtain 11-3 as ayellow oil (1.9 g, 100% yield). ¹H NMR (400 MHz, CDCl₃) δ 8.04 (s, 1H),7.57 (d, J=7.8 Hz, 1H), 7.37 (d, J=7.9 Hz, 1H), 7.23 (t, J=7.5 Hz, 1H),7.16 (t, J=7.4 Hz, 1H), 7.01 (br s, 1H), 3.48-3.39 (m, 4H), 3.15-3.05(m, 4H), 2.96-2.89 (m, 2H), 2.89-2.80 (m, 2H); ¹³C NMR (101 MHz, CDCl₃)δ 136.3, 127.1, 122.3, 122.2, 122.1, 119.6, 118.5, 113.0, 111.5, 54.2,49.4 (d, J=13.2 Hz), 47.9, 23.3; ¹⁹F NMR (376 MHz, CDCl₃) δ +57.9; R_(f)(7:3 hexane:EtOAc): 0.27; ESI-MS (m/z): 381 [MH]⁺.

2,2′-((Furan-2-ylmethyl)azanediyl)diethanesulfonyl fluoride (10-4).Reaction in CH₂Cl₂ (0.33 M) in the presence of 2 equiv ESF. The crudeproduct was purified by column chromatography (95:5 to 7:3 hexane:EtOAc)to give 11-4 as a yellow oil (1.6 g, 99% yield). ¹H NMR (400 MHz, CDCl₃)δ 7.41 (dd, J=1.9, 0.8 Hz, 1H), 6.37 (dd, J=3.2, 1.9 Hz, 1H), 6.28 (dd,J=3.2, 0.7 Hz, 1H), 3.81 (s, 2H), 3.53 (td, J=6.9, 3.6 Hz, 4H), 3.16(td, J=7.0, 1.2 Hz, 4H); ¹³C NMR (101 MHz, CDCl₃) δ 149.9, 143.1, 110.7,110.1, 49.6, 49.4 (d, J=11.1 Hz), 47.9; ¹⁹F NMR (376 MHz, CDCl₃) δ+57.9; R_(f) (7:3 hexane:EtOAc): 0.47; ESI-MS (m/z): 340 [MNa]⁺.

2-((4-(2-Oxopiperidin-1-yl)phenyl)amino)ethanesulfonyl fluoride (10-5).Reaction in DMF (0.38 g substrate, 2 mmol) and ESF (0.2 mL, 1.1 equiv).The reaction mixture was stirred at 50° C. overnight, monitored byLC-MS. After that time, reaction mixture was concentrated by rotaryevaporation and the crude product was purified by column chromatographyto give a beige solid (570 mg, 95% yield). mp 188-189° C. R_(f) (EtOAc):0.2. ¹H NMR (400 MHz, DMSO-d₆) δ 7.08 (d, J=8 Hz, 2H), 6.61 (d, J=8 Hz,2H), 5.96 (t, J=8 Hz, 1H), 4.07 (dd, J=12, 4 Hz, 2H), 3.59 (dd, J=12, 4Hz, 2H), 3.47 (t, J=4 Hz, 2H), 2.31 (t, J=4 Hz, 2H), 1.80 (m, 4H); ¹³CNMR (101 MHz, DMSO-d₆) δ 168.6, 145.5, 133.3 127.1, 112.1, 51.3, 49.7(d, J=10 Hz), 37.6, 32.5, 23.1, 21.0; ¹⁹F NMR (376 MHz, DMSO-d₆) δ+58.4; ESI-MS (m/z): 301 [MH]⁺.

2-(1-((1S,2S)-2-Hydroxycyclohexyl)-2-(4-methylbenzoyl)-hydrazinyl)ethanesulfonylfluoride (10-6). Reaction in THE with substrate (0.5 g, 2 mmol) and ESF(0.2 mL, 1.1 equiv), giving a white crystalline solid (mp 174-176° C.)in quantitative yield (0.712 g). ¹H NMR (400 MHz, CDCl₃) δ 7.67 (d,J=8.2 Hz, 2H), 7.27 (d, J=8.0 Hz, 3H), 7.18 (s, 1H), 3.81-3.71 (m, 1H),3.65-3.57 (m, 1H), 3.57-3.50 (m, 2H), 3.26 (ddd, J=11.0, 9.4, 4.6 Hz,1H), 2.66 (ddd, J=11.4, 9.3, 3.6 Hz, 1H), 2.42 (s, 3H), 2.10-2.02 (m,1H), 1.91 (d, J=10.2 Hz, 1H), 1.84-1.77 (m, 1H), 1.77-1.68 (m, 1H),1.40-1.13 (m, 4H); ¹³C NMR (101 MHz, CDCl₃) δ 168.4, 143.5, 129.7,129.1, 127.3, 72.0, 70.4, 49.9 (d, J=15.3 Hz), 49.8, 33.0, 25.0, 24.2,24.1, 21.7; ¹⁹F NMR (376 MHz, CDCl₃) δ +56.8; ESI-MS (m/z): 359 [MH]⁺.

4-(3-Carboxy-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinolin-7-yl)-1-(2-(fluorosulfonyl)ethyl)piperazin-1-iumchloride (10-7). A suspension of the starting material (HCl salt) in DMF(0.5 M in substrate) was treated with 4 equiv ESF and the reactionmixture was stirred at 50° C. overnight. The product was collected as awhite precipitate by filtration, washed with hexanes, and dried (4.2 g,88% yield). ¹H NMR (400 MHz, TFA-d₁) δ 9.35 (s, 1H), 8.28 (d, J=12 Hz,1H), 8.00 (d, J=4 Hz, 1H), 4.34-4.29 (m, 4H), 4.14-4.11 (m, 3H), 4.04(t, J=8 Hz, 2H), 3.91 (t, J=12 Hz, 2H), 3.69 (t, J=12 Hz, 2H), 1.69 (d,J=8 Hz, 2H), 1.44 (d, J=4 Hz, 2H); ¹³C NMR (151 MHz, DMSO-d₆ with Py-d₅)δ 176.5, 166.1, 153.1 (d, J=250.0 Hz), 147.8, 145.0 (d, J=10.1 Hz),139.0, 118.8 (d, J=7.8 Hz), 111.0 (d, J=22.7 Hz), 106.9, 106.3 (d, J=3.2Hz), 51.7, 50.9, 49.2 (d, J=4.6 Hz), 48.1 (d, J=11.5 Hz), 35.7, 7.5; ¹⁹FNMR (376 MHz, DMSO-d₆ with Py-d₅) 6±59.3, −121.4; ESI-MS (m/z): 442[MH]⁺.

2-(4-(2-chlorodibenzo[b,f][1,4]oxazepin-11-yl)piperazin-1-yl)ethanesulfonylfluoride (10-8). Reaction in CH₂Cl₂ with the starting amine (314 mg, 1mmol) and ESF (0.1 mL, 1.1 equiv), giving the desired product as a whitesolid (420 mg, quant. yield). mp 158-159° C. ¹H NMR (400 MHz,Acetone-d₆) δ 7.57 (dd, J=8, 4 Hz, 1H), 7.46 (d, J=4 Hz, 1H), 7.37 (d,J=8 Hz, 1H), 7.17-7.15 (m, 1H), 7.12-7.07 (m, 2H), 7.03-6.98 (m, 1H),4.03 (m, 2H), 3.57 (m, 4H), 3.07 (m, 2H), 2.79 (m, 4H); ¹³C NMR (101MHz, CDCl₃) δ 159.3, 158.7, 151.9, 139.8, 134.4, 132.8, 130.3, 129.0,127.1, 125.9, 124.9, 124.8, 122.8, 120.2, 52.4, 51.2, 48.7 (d, J=15 Hz),47.2; ¹⁹F NMR (376 MHz, CDCl₃) δ +57.5; ESI-MS (m/z): 424 [MH]⁺.

2-(2-Methyl-4-nitro-1H-imidazol-1-yl)ethanesulfonyl fluoride (10-9).Reaction in THE with starting imidazole (38 mg, 0.3 mmol) and ESF (0.03mL; 1.1 equiv), giving a white crystalline solid. Further purificationwas performed by flash column chromatography to provide the desiredproduct (65 mg, 91% yield). mp 164-165° C. R_(f) (EtOAc): 0.60; ¹H NMR(400 MHz, Acetone-d₆) δ 8.23 (s, 1H), 4.81 (t, J=8 Hz, 2H), 4.53 (q, J=8Hz, 2H), 2.50 (s, 3H); ¹³C NMR (101 MHz, acetone-d₆) δ 206.3, 146.3,121.7, 50.6 (d, J=16.2 Hz), 41.7, 12.9; ¹⁹F NMR (376 MHz, Acetone-d₆) δ+56.8; ESI-MS (m/z): 238 [MH]⁺.

Section C: General procedure for the reaction of ESF with sulfonamidesand alcohols. The starting material (1 equiv) and triphenylphosphine(0.1 equiv) were dissolved in CH₂Cl₂ (0.33 M in substrate) and treatedwith ESF (1-2.5 equiv). The reaction mixture was stirred at roomtemperature overnight, monitoring conversion by LC-MS, GC-MS, or TLC asappropriate. Upon completion, CH₂Cl₂ and excess of ESF were removed byrotary evaporation and the crude product was purified by chromatographyon a short silica gel column.

2-(4-Methyl-N-(prop-2-yn-1-yl)phenylsulfonamido)ethanesulfonyl fluoride(10-10) was obtained as a white solid (251 mg, 78% yield). R_(f) (5:1hexane:EtOAc): 0.25; mp 125-126° C.; ¹H NMR (400 MHz, CDCl₃) δ 7.74 (d,J=8.3 Hz, 2H), 7.35 (d, J=8.5 Hz, 2H), 4.15 (d, J=2.5 Hz, 2H), 3.87-3.80(m, 2H), 3.71-3.65 (m, 2H), 2.45 (s, 3H), 2.19 (t, J=2.5 Hz, 1H); ¹³CNMR (101 MHz, CDCl₃) δ 144.8, 134.5, 130.1, 127.9, 76.2, 75.0, 50.4 (d,J=16.0 Hz), 41.9, 38.8, 21.8; ¹⁹F NMR (376 MHz, CDCl₃) δ 55.9; ESI-MS(m/z): 320 [MH]⁺.

2,2′-(((4-(3-Phenyl-5-(trifluoromethyl)-1H-pyrazol-1-yl)phenyl)sulfonyl)azanediyl)-diethanesulfonylfluoride (10-11) was obtained as a white solid (175 mg, 60% yield).R_(f) (5:1 hexane:EtOAc): 0.21; mp 135-137° C.; ¹H NMR (400 MHz, CDCl₃)δ 7.84 (d, J=8.6 Hz, 2H), 7.58 (d, J=8.6 Hz, 2H), 7.47-7.43 (m, 1H),7.43-7.39 (m, 1H), 7.28-7.23 (m, 3H), 6.80 (s, 1H), 3.86-3.80 (m, 4H),3.71-3.66 (m, 4H); ¹³C NMR (101 MHz, CDCl₃) δ 145.4, 144.8 (d, J=38.3Hz), 143.9, 135.7, 130.0, 129.4, 129.0, 128.8, 128.6, 117.4 (d, J=336.3Hz), 107.2, 50.7, 50.6 (d, J=16.1 Hz), 45.6; ¹⁹F NMR (376 MHz, CDCl₃) δ+59.4, −62.8; ESI-MS (m/z): 588 [MH]⁺.

2-(Prop-2-yn-1-yloxy)ethanesulfonyl fluoride (10-12) was isolated as acolorless oil (2 g, 60% yield). Since the product is quite volatile,evaporative removal of EtOAc was done gently, leaving a small amount ofthe solvent in the sample. R_(f) (7:3 hexane:EtOAc): 0.43. ¹H NMR (400MHz, CDCl₃) δ 4.20 (d, J=4 Hz, 2H), 4.00 (dt, J=4, 8 Hz, 2H), 3.66 (m,2H), 2.51 (t, J=4 Hz, 1H); ¹³C NMR (101 MHz, CDCl₃) δ 78.3, 75.9, 62.5,58.6, 51.0 (d, J=17.2 Hz); ¹⁹F NMR (376 MHz, CDCl₃) δ +58.5. EI-MS(m/z): 166 [M]⁺.

Section D: 2,2′-((3-Ethynylphenyl)azanediyl)diethanesulfonyl fluoride(10-13). (adapted from Hyatt et al. J. Org. Chem., 1979, 44, 3847-3858)ESF (1.8 mL, 20 mmol) was added to aniline (1.17 g, 10 mmol) in glacialacetic acid (3 mL) and the reaction mixture was stirred at 50° C. for 24h. Upon completion, the crude product was isolated by filtration, washedwith hexanes, and recrystallized from CCl₄—CH₂Cl₂. The desired productwas obtained as light brown crystals (2.94 g, 87% yield). mp 98-100° C.¹H NMR (400 MHz, CDCl₃) δ 7.31 (t, J=8.0 Hz, 1H), 7.10-7.06 (m, 1H),6.85-6.82 (m, 1H), 6.78-6.73 (m, 1H), 4.01 (t, J=6.4 Hz, 4H), 3.67-3.59(m, 4H), 3.10 (s, 1H); ¹³C NMR (101 MHz, CDCl₃) δ 144.0, 130.5, 124.3,124.2, 117.6, 115.0, 83.3, 78.0, 48.3 (d, J=14.4 Hz), 46.8; ¹⁹F NMR (376MHz, CDCl₃) δ +57.2; ESI-MS (m/z): 338 [MH]⁺.

Section E: 2-(Naphthalen-2-yloxy)ethanesulfonyl fluoride (10-14).2-Naphthol (0.29 g; 2 mmol) dissolved in THE (2 mL) was treated with ESF(0.2 mL, 1.1 mmol), followed by addition of tetrabutylammonium fluoride(0.2 mL of 1M solution in THF, 10 mol %). The reaction mixture wasstirred at room temperature for 48 h. After removal of solvent by rotaryevaporation, the crude product was purified by column chromatography(95:5 hexane:EtOAc) to give the product as a white solid (0.42 g, 81%yield). mp 70-72° C. ¹H NMR (400 MHz, CDCl₃) δ 7.82-7.74 (m, 3H), 7.50(t, J=8 Hz, 1H), 7.41 (t, J=8 Hz, 1H) 7.19-7.13 (m, 2H), 4.54 (m, 2H);¹³C NMR (101 MHz, CDCl₃) δ 155.3 134.2, 130.0, 129.6, 127.8, 126.9,126.8, 124.4, 118.5, 107.3, 61.0, 50.6 (d, J=17.1 Hz); ¹⁹F NMR (376 MHz,CDCl₃) δ +59.1; GC-MS (t_(R)): 6.56 min; EI-MS (m/z): 254 [M]⁺.

Section F: General procedure for the reaction of ESF with 1,3-dicarbonylcompounds. The starting compound (1 equiv) and quinine (0.1 equiv) weredissolved in CH₂Cl₂ (0.33M in substrate) and treated with ESF (1.1equiv). The reaction mixture was stirred at room temperature for severalhours, monitoring conversion by LC-MS, GC-MS, or TLC, as appropriate.Upon completion, CH₂Cl₂ and excess of ESF were removed by rotaryevaporation and the crude product was purified by short columnchromatography (9:1 to 6:4 hexane:EtOAc).

2-(1-Acetyl-2-oxocyclopentyl)ethanesulfonyl fluoride (10-15) wasobtained as a white solid (2.23 g, 95% yield). mp 76-78° C. ¹H NMR (400MHz, CDCl₃) δ 3.47-3.31 (m, 4H), 2.60-2.47 (m, 1H), 2.43 (br s, 2H),2.34-2.24 (m, 2H), 2.15 (s, 3H), 2.07-1.96 (m, 4H), 1.81 (dt, J=12.6,6.5 Hz, 1H); ¹³C NMR (101 MHz, CDCl₃) δ 215.3, 203.5, 65.7, 46.7 (d,J=18.2 Hz), 38.4, 32.6, 26.9, 26.5, 19.7; ¹⁹F NMR (376 MHz, CDCl₃) δ+51.8; R_(f) (7:3 hexane:EtOAc): 0.29; EI-MS (m/z): 194 [M-COMe]⁺, 236[M]⁺.

Ethyl 1-(2-(fluorosulfonyl)ethyl)-2-oxocyclopentane-carboxylate (10-16)was obtained as a white solid (2.4 g, 88% yield). mp 36-38.5° C. ¹H NMR(400 MHz, CDCl₃) δ 4.17 (qq, J=7.2, 3.6 Hz, 2H), 3.87-3.73 (m, 1H),3.57-3.43 (m, 1H), 2.56-2.42 (m, 2H), 2.41-2.27 (m, 1H), 2.30-2.17 (m,2H), 2.15-2.03 (m, 1H), 2.03-1.86 (m, 2H), 1.23 (t, J=7.1 Hz, 3H); ¹³CNMR (101 MHz, CDCl₃) δ 213.9, 170.6, 62.2, 57.5, 46.9 (d, J=17.6 Hz),38.0, 34.9, 27.3, 19.7, 14.1; ¹⁹F NMR (376 MHz, CDCl₃) δ +51.4; R_(f)(7:3 hexane:EtOAc): 0.50; EI-MS (m/z): 266 [M]⁺.

3-Cyano-4-oxo-4-(piperidin-1-yl)butane-1-sulfonylfluoride (10-17) wasobtained as thick yellow oil (2.0 g, 76% yield). ¹H NMR (400 MHz, CDCl₃)δ 3.96 (t, J=8 Hz, 1H), 3.81-3.68 (m, 2H), 3.65-3.51 (m, 2H), 3.49-3.39(m, 2H), 2.60 (m, 2H), 1.80-1.58 (m, 6H); ¹³C NMR (101 MHz, CDCl₃) δ160.5, 115.8, 47.8 (d, J=18.1 Hz), 47.1, 44.0, 31.8, 25.7, 25.2, 23.9,23.4; ¹⁹F NMR (376 MHz, CDCl₃) δ +53.7; R_(f) (7:3 hexane:EtOAc): 0.29;EI-MS (m/z): 262 [M]⁺.

Ex. 1(J). Aryl Fluorosulfates Prepared by a Convenient Procedure withGaseous SO₂F₂

Section A. Representative procedure using triethylamine as base:propane-2,2-diylbis(4,1-phenylene) difluorosulfonate (12-10). A 2-litersingle-neck round-bottom flask was charged with bisphenol-A (114.9 g,0.5 mol), CH₂Cl₂ (1 L) and Et₃N (174 mL, 1.25 mol, 2.5 equiv). Themixture was stirred at room temperature for 10 min. The reaction flaskwas then sealed with a septum, the atmosphere above the solution wasremoved with gentle vacuum, and SO₂F₂ gas (sulfuryl fluoride, VIKANE)was introduced by needle from a balloon filled with the gas. For largescale reactions such as this, depletion of the sulfuryl fluoride fromthe balloon was easily observed, and more reagent was introduced with afresh balloon when required. For small scale reactions, SO₂F₂ was usedin excess. These reactions can be easily followed by TLC.

The reaction mixture was vigorously stirred at room temperature for 2-4hours, monitoring by GC-MS and TLC. After completion, the solvent wasremoved by rotary evaporation, the residue was dissolved in EtOAc (1 L),and the solution was washed with 1N HCl (2×500 mL) and brine (2×500 mL).The organic phase was dried over anhydrous Na₂SO₄ and concentrated. Theresulting solid was dried under high vacuum at 60° C. overnight to givethe desired compound as a white crystalline solid in quantitative yield(197.1 g, 100% yield). mp 48-49° C. ¹H NMR (400 MHz, CDCl₃) δ 7.34-7.32(m, 2H), 7.28-7.26 (m, 2H), 1.72 (s, 3H); ¹³C NMR (101 MHz, CDCl₃) δ150.4, 148.2, 128.7, 120.5, 42.9, 28.4, 30.7; ¹⁹F NMR (376 MHz, CDCl₃) δ+37.0; GC-MS (t_(R)): 7.2 min; EI-MS (m/z): 392 [M]⁺.

Phenyl fluorosulfonate (12-1) was isolated as a colorless oil in 94%yield (1.65 g). ¹H NMR (400 MHz, CDCl₃) δ 7.51-7.47 (m, 2H), 7.43-7.41(m, 1H), 7.36-7.34 (2H); ¹³C NMR (101 MHz, CDCl₃) δ 150.1, 130.4, 128.6,120.8; ¹⁹F NMR (376 MHz, CDCl₃) δ +37.0; GC-MS (t_(R)): 3.85 min; EI-MS(m/z): 176 [M]⁺.

3-Acetamidophenyl fluorosulfonate (12-2) was isolated as a light brownsolid (mp 113-114° C.) in 99% yield (2.3 g). ¹H NMR (400 MHz, CDCl3) δ8.45 (bs, 1H), 7.80 (s, 1H), 7.44-7.41 (m, 1H), 7.38-7.34 (m, 1H),7.06-7.04 (m, 1H), 2.19 (s, 3H); ¹³C NMR (101 MHz, CDCl3) δ 169.4,150.1, 139.9, 130.4, 119.5, 116.1, 112.5, 24.3; ¹⁹F NMR (376 MHz, CDCl3)δ +37.4; GC-MS (t_(R)): 6.0 min; EI-MS (m/z): 233 [M]⁺.

(1,1′-Biphenyl)-4-yl fluorosulfonate (12-3) was isolated as a whitesolid (mp 89-90° C.) in 99% yield (5.0 g). ¹H NMR: (400 MHz, CDCl₃) δ7.70-7.66 (m, 2H), 7.60-7.57 (m, 2H), 7.52-7.47 (m, 2H), 7.45-7.41 (m,3H); ¹³C NMR: (101 MHz, CDCl₃) δ 149.4, 141.9, 139.2, 129.0, 128.1,127.2, 121.1; ¹⁹F NMR (376 MHz, CDCl₃) δ +37.1; GC-MS (t_(R)): 5.9 min;EI-MS (m/z): 252 [M]⁺.

4-Nitrophenyl fluorosulfonate (12-4) was isolated as yellow oil in 95%yield (4.2 g). ¹H NMR (400 MHz, CDCl₃) δ 8.37 (dd, J=7.2 Hz, 2.4 Hz,2H), 7.55 (m, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 153.3, 147.3, 126.1,122.1; ¹⁹F NMR (376 MHz, CDCl₃) δ +38.9; GC-MS (t_(R)): 4.9 min; EI-MS(m/z): 221 [M]⁺.

4-Aminophenyl fluorosulfonate (12-5) was isolated as brown solid (mp41-42° C.) in 91% yield (8.0 g) according to the general procedure F17using 3 equiv of Et₃N. ¹HNMR (400 MHz, CDCl₃) δ 7.08 (d, J=8.5 Hz, 1H),6.65 (d, J=8.7 Hz, 1H), 3.87 (br s, 1H); ¹³C NMR (101 MHz, CDCl₃) δ146.9, 142.1, 121.8, 115.6, 115.5; ¹⁹F NMR (376 MHz, CDCl₃) δ +35.5;GC-MS (t_(R)): 5.0 min; EI-MS (m/z): 191 [M]⁺.

2-Isopropyl-5-methylphenyl fluorosulfonate (12-6) was isolated as acolorless oil in 99% yield (2.4 g). ¹H NMR (400 MHz, CDCl₃) δ 7.29 (d,J=8 Hz, 1H), 7.18 (d, J=8 Hz, 1H), 7.13 (s, 1H), 3.28 (q, J=2.8 Hz, 1H),2.37 (s, 3H), 1.27 (d, J=2.4 Hz, 3H), 1.25 (d, J=2.4 Hz, 3H); ¹³C NMR(101 MHz, CDCl₃) δ 147.8, 137.8, 137.4, 129.6, 127.6, 120.9, 26.7, 23.1,20.8; ¹⁹F NMR (376 MHz, CDCl₃) δ +38.9; GC-MS (t_(R)): 4.6 min; EI-MS(m/z): 232 [M]⁺.

3-Methoxyphenyl fluorosulfonate (12-7) was isolated as a colorless oilin 91% yield (0.4 g). ¹H NMR (400 MHz, CDCl₃) δ 7.36 (m, 1H), 6.96-6.92(m, 2H), 6.86 (m, 1H), 3.83 (s, 3H); ¹³C NMR (101 MHz, CDCl₃) δ 161.0,150.8, 130.8, 114.5, 112.7, 107.0, 55.8; ¹⁹F NMR (376 MHz, CDCl₃) δ+37.2; GC-MS (t_(R)): 4.5 min; EI-MS (m/z): 206 [M]⁺.

Naphthalen-2-yl fluorosulfonate (12-8) was isolated as an off-whitesolid (mp 34-35° C.) in 98% yield (22.13 g). ¹H NMR (400 MHz, CDCl₃) δ7.95 (d, J=9.1 Hz, 1H), 7.93-7.87 (m, 2H), 7.82 (d, J=2.6 Hz, 1H),7.64-7.54 (m, 2H), 7.44 (ddd, J=9.0, 2.5, 0.9 Hz, 1H); ¹³C NMR (101 MHz,CDCl₃) δ 147.7, 133.5, 132.6, 131.0, 128.2, 128.1, 127.8, 127.5, 119.1,119.0; ¹⁹F NMR (376 MHz, CDCl₃) δ 37.2; GC (t_(R)): 5.4 min; EI-MS(m/z): 226 [M]⁺.

1,4-Phenylene difluorosulfonate (12-9) was isolated as a light brownsolid (mp 92-93° C.) in 92% yield (5.1 g). ¹H NMR (400 MHz, CDCl₃) δ7.21 (s, 4H); ¹³C NMR (101 MHz, CDCl₃) δ 142.6, 125.3, 111.7; ¹⁹F NMR(376 MHz, CDCl₃) δ +40.6; GC-MS (t_(R)): 4.7 min; EI-MS (m/z): 274 [M]⁺.

(E)-Hex-3-ene-3,4-diylbis(4,1-phenylene) difluorosulfonate (12-11) wasisolated as a white solid (mp 122-123° C.) in 95% yield (0.8 g). ¹H NMR(400 MHz, CDCl₃) δ 7.38-7.36 (m, 2H), 7.33-7.30 (m, 2H), 2.12 (q, J=7.6Hz, 2H), 0.78 (t, J=7.6 Hz, 3H); ¹³C NMR (101 MHz, CDCl₃) δ 148.7,142.6, 138.7, 130.5, 120.6, 28.4, 13.1; ¹⁹F NMR (376 MHz, CDCl₃) δ+37.0; GC-MS (t_(R)): 7.5 min; EI-MS (m/z): 432 [M]⁺.

(E)-2-((2-Fluorosulfoxybenzylidene)amino)benzene fluorosulfate (12-12)was isolated as a grey solid (mp 78-79° C.) in 95% yield (1.8 g). ¹H NMR(400 MHz, CDCl₃) δ 8.79 (s, 1H), 8.39 (dd, J=8.0, 2.0 Hz, 1H), 7.66-7.62(m, 1H), 7.57-7.53 (m, 1H), 7.49-7.37 (m, 2H), 7.35-7.26 (m, 1H), 7.25(dd, J=8.0, 2 Hz, 1H); ¹³C NMR (100 MHz, CDCl₃) δ 155.3, 149.8, 143.6,143.4, 133.7, 129.7, 129.6, 129.2, 127.9, 122.1, 121.5, 120.2; ¹⁹F NMR(375 MHz, CDCl₃) δ 40.0, 38.4; GC-MS (t_(R)): 6.9 min; EI-MS (m/z): 377[M]⁺.

(3-Oxo-1,3-dihydroisobenzofuran-1,1-diyl)bis(4,1-phenylene)difluorosulfonate (12-13) was isolated as a white solid (mp 89-91° C.)in 94% yield (11.3 g). ¹H NMR: (400 MHz, CDCl₃) δ 7.98 (d, J=8.0 Hz,1H), 7.79 (m, 1H), 7.66-7.63 (m, 1H), 7.59 (d, J=7.6 Hz, 1H), 7.49-7.45(m, 4H), 7.36-7.33 (m, 4H); ¹³C NMR (101 MHz, CDCl₃) δ 168.5, 150.2,149.9, 140.9, 134.8, 130.2, 129.2, 126.6, 125.2, 123.8, 121.3, 89.5; ¹⁹FNMR (376 MHz, CDCl₃) δ +37.7; GC-MS (t_(R)): 9.9 min; EI-MS (m/z): 482[M]⁺.

Ethane-1,1,1-triyltris(benzene-4,1-diyl) trifluorosulfonate (12-14) wasisolated as a white solid (mp 94-95° C.) in quantitative yield (5.6 g).¹H NMR (400 MHz, CDCl₃) δ 7.33-7.30 (m, 6H), 7.21-7.18 (m, 6H), 2.23 (s,3H); ¹³C NMR (101 MHz, CDCl₃) δ 148.6, 148.0, 130.4, 120.8, 52.0, 30.8;¹⁹F NMR: (376 MHz, CDCl₃) δ +37.4; GC-MS (t_(R)): 9.3 min; EI-MS (m/z):454 [M-OSO2F]⁺.

(8R,9S,13S,14S,17R)-17-Ethynyl-17-hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-3-ylfluorosulfonate (12-15) was isolated as a thick colorless gel in 96%yield (0.8 g). ¹H NMR (400 MHz, CDCl₃) δ 7.36 (d, J=8.4 Hz, 1H), 7.09(dd, J=8.8 Hz, 2.4 Hz 1H), 7.03 (d, J=2.0 Hz, 1H), 2.90-2.87 (m, 2H),2.62 (s, 1H), 2.38-2.25 (m, 3H), 2.17 (s, 1H), 2.05 (m, 1H), 1.94-1.90(m, 2H), 1.81-1.68 (m, 3H), 1.53-1.36 (m, 4H), 0.89 (s, 3H); ¹³C NMR(101 MHz, CDCl₃) δ 147.9, 140.9, 139.5, 127.2, 120.5, 117.5, 87.3, 79.6,74.1, 49.3, 46.9, 43.6, 38.8, 38.6, 32.5, 29.4, 26.6, 26.1, 22.7, 12.5;¹⁹F NMR (376 MHz, CDCl₃) δ +36.8; GC-MS (t_(R)): 8.5 min; EI-MS (m/z):378 [M]⁺.

3-(Diphenylphosphoryl)phenyl fluorosulfonate (12-16) was isolated as athick yellow oil in 97% yield (3.7 g). ¹H NMR (400 MHz, CDCl₃) δ7.72-7.65 (m, 2H), 7.65-7.57 (m, 8H), 7.57-7.51 (m, 5H), 7.51-7.48 (m,1H), 7.48-7.41 (m, 8H); ¹³C NMR (101 MHz, CDCl₃) δ 145.0, 149.8, 136.9,135.9, 132.6, 132.60, 132.2, 132.1, 132.0, 131.9, 131.5, 131.0, 130.9,130.5, 128.9, 128.8, 124.5 (d, J=5.3 Hz), 124.4 (d, J=3.2 Hz); ¹⁹F NMR(376 MHz, CDCl₃) δ 38.0; R_(f) (EtOAc): 0.6; ESI-MS (m/z): 377 [MH]⁺.

5-Chloroquinolin-8-yl fluorosulfonate (12-17) was isolated as a paleyellow solid (mp 105-107° C.) in 99% yield (6.5 g) following generalprocedure F17 with 2 equivalents of DIPEA. ¹H NMR (400 MHz, CDCl₃) δ9.09 (dd, J=4.1, 1.6 Hz, 1H), 8.60 (dd, J=8.6, 1.6 Hz, 1H), 7.72-7.61(m, 3H); ¹³C NMR (101 MHz, CDCl₃) δ 152.4, 144.8, 141.0, 133.4, 132.1,127.9, 125.9, 123.6, 121.4; ¹⁹F NMR (376 MHz, CDCl₃) δ 40.5; R_(f) (7:3hexane:EtOAc): 0.67; GC (t_(R)): 5.98 min; EI-MS (m/z): 261 [M]⁺.

2-Oxo-2H-chromen-7-yl fluorosulfonate (12-18) was isolated as anoff-white solid (mp 121-124° C.) in 98% yield (11.9 g). ¹H NMR (400 MHz,CDCl₃) δ 7.73 (dd, J=9.7, 0.6 Hz, 1H), 7.62 (d, J=8.6 Hz, 1H), 7.35 (d,J=2.3 Hz, 1H), 7.29 (ddd, J=8.5, 2.4, 0.9 Hz, 1H), 6.50 (d, J=9.7 Hz,1H); ¹³C NMR (101 MHz, CDCl₃) δ 159.3, 154.8, 151.3, 142.2, 129.8,119.1, 118.1, 117.3, 110.4; ¹⁹F NMR (376 MHz, CDCl₃) δ 38.8; GC (t_(R)):5.94 min; EI-MS (m/z): 244 [M]⁺.

4-(2-Amino-2-oxoethyl)phenyl fluorosulfonate (12-19) was isolated as awhite solid (mp 109-110° C.) in 96% yield (11.22 g). ¹H NMR (400 MHz,CDCl₃) δ 7.38 (d, J=8.6 Hz, 2H), 7.31 (d, J=8.4 Hz, 2H), 6.18 (br s,1H), 5.70 (br s, 1H), 3.58 (s, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 172.9,149.3, 135.7, 131.5, 121.4, 42.2; ¹⁹F NMR (376 MHz, CDCl₃) δ 37.2;ESI-MS (m/z): 234 [MH]⁺.

2-(2H-Benzo[d][1,2,3]triazol-2-yl)-4-methylphenyl fluorosulfonate(12-20) was isolated as a thick colorless oil (solidifies upon standing)was isolated in 98% yield (4.54 g) following the general procedure using4 equivalents of triethylamine. Further purification was performed bysilica gel column chromatography (5-20% EtOAc in hexane). mp 63-64° C.¹H NMR (400 MHz, CDCl₃) δ 8.04 (d, J=2.1 Hz, 1H), 7.98-7.92 (m, 2H),7.50-7.42 (m, 3H), 7.35 (dd, J=8.5, 2.2 Hz, 1H), 2.49 (s, 3H); ¹³C NMR(101 MHz, CDCl₃) δ 145.3, 140.4, 139.6, 132.5, 131.2, 127.9, 127.0,123.0, 118.7, 21.1; ¹⁹F NMR (376 MHz, CDCl₃) δ 42.4; R_(f) (9:1hexane:EtOAc): 0.44; ESI-MS (m/z): 308 [MH]⁺.

Benzo[d][1,3]dioxol-5-yl fluorosulfonate (12-21) was isolated as acolorless oil in 88% yield (9.68 g). ¹H NMR (400 MHz, CDCl₃) δ 6.83-6.80(m, 3H), 6.05 (s, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 148.7, 147.8, 144.2,114.2, 108.4, 103.1, 102.7; ¹⁹F NMR (376 MHz, CDCl₃) δ 36.1; R_(f) (9:1hexane:EtOAc): 0.54; GC (t_(R)): 4.82 min; EI-MS (m/z): 220 [M]⁺.

6-Methylpyridin-3-yl fluorosulfonate (12-22) was isolated as colorlessneedles in 78% yield (2.97 g). Crude product was purified by extractionwith EtOAc (50 mL×2); wash with NaHCO₃ (35 mL), brine (10 mL), followedby filtration through short column (SiO2; 30% EtOAc in hexane). mp33-34.5° C.; ¹H NMR (400 MHz, CDCl₃) δ 8.52 (d, J=2.9 Hz, 1H), 7.57 (dd,J=8.6, 2.6 Hz, 2H), 7.27 (d, J=8.6 Hz, 1H), 2.60 (s, 3H); ¹³C NMR (101MHz, CDCl₃) δ 159.5, 145.4, 141.8, 128.8, 124.5, 24.1; ¹⁹F NMR (376 MHz,CDCl₃) δ 37.4; R_(f) (7:3 hexane:EtOAc): 0.46; ESI-MS (m/z): 192 [MH]⁺.

5-Formyl-2-methoxyphenyl fluorosulfonate (12-23) was isolated as yellowoil in 98% yield (4.25 g). Crude product was purified by flash columnchromatography (SiO2; 30% EtOAc in hexane). ¹H NMR (400 MHz, CDCl₃) δ9.84 (s, 1H), 7.87 (dd, J=8.5, 2.0 Hz, 1H), 7.81 (dd, J=2.0, 1.2 Hz,1H), 7.18 (d, J=8.5 Hz, 1H), 3.98 (s, 3H); ¹³C NMR (101 MHz, CDCl₃) δ189.1, 156.0, 139.2, 132.4, 129.9, 122.7, 113.4, 56.7; ¹⁹F NMR (376 MHz,CDCl₃) δ 40.1; R_(f) (7:3 hexane:EtOAc): 0.41; GC (t_(R)): 5.4 min;EI-MS (m/z): 234 [M]⁺.

4-Formyl-2-methoxyphenyl fluorosulfonate (12-24) was isolated as yellowsolid (mp 45-49° C.) in 97% yield (45.37 g). ¹H NMR (400 MHz, CDCl₃) δ9.94 (s, 1H), 7.70-7.34 (m, 3H), 3.96 (s, 3H); ¹³C NMR (101 MHz, CDCl₃)δ 190.4, 152.0, 142.7, 137.1, 123.9, 123.2, 112.2, 56.5; ¹⁹F NMR (376MHz, CDCl₃) δ 40.4; R_(f) (9:1 hexane:EtOAc): 0.36; GC (t_(R)): 5.2 min;EI-MS (m/z): 234 [M]⁺.

4-Allyl-2-methoxyphenyl fluorosulfonate (12-25) was isolated as acolorless oil in 97% yield (119.56 g). Crude product was purified byfiltration through a short pad of silica (10% EtOAc in hexane) to removebrown impurities. ¹H NMR (400 MHz, CDCl₃) δ 7.23 (dd, J=8.3, 1.3 Hz,1H), 6.90 (d, J=2.0 Hz, 1H), 6.83 (ddt, J=8.3, 2.0, 0.6 Hz, 1H),6.02-5.91 (m, 1H), 5.16 (td, J=1.5, 0.9 Hz, 1H), 5.15-5.11 (m, 1H), 3.90(s, 3H), 3.43-3.40 (m, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 151.0, 142.4,137.4, 136.3, 122.1, 120.9, 116.9, 113.7, 56.1, 40.1; ¹⁹F NMR (376 MHz,CDCl₃) δ 38.9; R_(f) (9:1 hexane:EtOAc): 0.64; GC (t_(R)): 5.3 min;EI-MS (m/z): 246 [M]⁺.

Mesityl fluorosulfonate (12-26) was prepared using 4 equivtriethylamine, followed by short filtration through a plug of silica gelto give the desired product in 85% yield (0.68 g) as a colorless liquid.¹H NMR (400 MHz, CDCl₃) δ 6.97-6.94 (m, 2H), 2.39-2.36 (m, 6H), 2.32 (s,3H); ¹³C NMR (101 MHz, CDCl₃) δ 146.9, 138.2, 130.5, 129.2, 20.7, 16.5;¹⁹F NMR (376 MHz, CDCl₃) δ 42.4; R_(f) (95:5 hexane:EtOAc): 0.72; GC(t_(R)): 4.6 min; EI-MS (m/z): 218 [M]⁺.

Catechol cyclic sulfate (12-27) was obtained as a colorless crystallinesolid (1.6 g, 92% yield). mp 33-35.5° C. ¹H NMR (400 MHz, CDCl₃) δ7.23-7.18 (m, 4H); ¹³C NMR (101 MHz, CDCl₃) δ 142.6, 125.3, 111.7; GC-MS(t_(R)): 4.4 min; EI-MS (m/z): 172 [M]⁺.

7,7,7′,7′-Tetramethyl-6,6′,7,7′-tetrahydro-5,5′-spirobi[indeno[5,6-d][1,3,2]dioxathiole]2,2,2′,2′-tetraoxide (12-28). The above procedure was employed with 6equiv Et₃N in CH₂Cl₂ solvent (0.25 M in substrate). The crude productwas suspended in a small amount of hexane (dissolving a brown impurity)and 12-28 was isolated by filtration as a white crystalline solid (15.27g). A second batch of the product was obtained after concentration ofthe mother liquor (3.18 g), giving a total of (18.55 g, 80% yield). mp223-225° C. R_(f) (9:1 hexane:EtOAc) 0.54. ¹H NMR (400 MHz, CDCl₃) δ7.02 (s, 2H), 6.62 (s, 2H), 2.32 (dd, J=90.4, 13.3 Hz, 4H), 1.41 (s,6H), 1.35 (s, 6H); ¹³C NMR (101 MHz, CDCl₃) δ 149.45, 146.07, 142.40,142.14, 107.79, 106.06, 58.81, 58.31, 44.24, 31.70, 30.20.

3-(o-Tolyl)naphtho[2,3-e][1,2,3]oxathiazin-4(3H)-one 2,2-dioxide(12-29). The above procedure was employed with 3.5 equiv Et₃N. The crudeproduct was suspended in a small amount of CH₂Cl₂ and filtered. The pureproduct was collected as a white powder and dried (2.81 g, 78% yield).mp 203-204° C. ¹H NMR (400 MHz, CDCl₃) δ 8.81 (s, 1H), 8.08-8.05 (m,1H), 7.96-7.93 (m, 1H), 7.81 (s, 1H), 7.74 (ddd, J=8.3, 6.9, 1.3 Hz,1H), 7.64 (ddd, J=8.2, 6.9, 1.2 Hz, 1H), 7.48-7.34 (m, 4H), 2.36 (s,3H); ¹³C NMR (101 MHz, CDCl₃) δ 160.4, 145.9, 138.5, 136.7, 133.8,131.9, 131.1, 131.0, 130.9, 130.7, 130.0, 129.8, 127.9, 127.7, 127.6,116.1, 116.0, 18.1; EI-MS (m/z): 339 [M]⁺.

II. Representative procedure for biphasic reaction conditions.3,5-Bis(fluorosulfonate)benzoic acid (12-30). 3,5-Dihydroxybenzoic acid(0.77 g, 5 mmol) was dissolved in a 3:2 CH₂Cl₂:water mixture (0.5 M).Diisopropylethylamine (4.5 mL, 25 mmol) was added, the reaction flaskwas sealed with a septum, air was evacuated, and a balloon filled withSO₂F₂ gas was introduced. The reaction mixture was vigorously stirredunder SO₂F₂ atmosphere at room temperature overnight. Upon completion,the volatiles were removed by rotary evaporation, and the residue wasacidified and extracted with CH₂Cl₂. The desired product was obtained asa white solid (1.45 g, 91% yield). mp 127-128° C. ¹H NMR (400 MHz,CDCl₃) δ 10.8 (br s, 1H), 8.19 (d, J=2.3 Hz, 2H), 7.68 (t, J=2.3 Hz,1H); ¹³C NMR (101 MHz, CDCl₃) δ 168.1, 150.1, 133.8, 123.3, 120.3; ¹⁹FNMR (376 MHz, CDCl₃) δ +39.1; ESI-MS (m/z): 317 [M]⁺.

(4R,4aS,7aR,12bS)-3-(Cyclopropylmethyl)-4a-hydroxy-7-oxo-2,3,4,4a,5,6,7,7a-octahydro-1H-4,12-methanobenzofuro[3,2-e]isoquinolin-9-ylfluorosulfate (12-31). The above procedure was performed starting withNaltrexone-HCl salt dihydrate (50 mg, 0.12 mmol) and 3 equiv Et₃N in 1:1CH₂Cl₂:water (0.1 M). After workup (sat. NaHCO₃) and purification bycolumn chromatography (95:5 CH₂Cl₂:MeOH, R_(f) 0.47), the desiredproduct was obtained as a white solid (42 mg, 82% yield). mp 135-138° C.¹H NMR (600 MHz, CDCl₃) δ 7.06 (d, J=8.4 Hz, 1H), 6.75 (d, J=8.4 Hz,1H), 4.80 (s, 1H), 3.23 (d, J=5.8 Hz, 1H), 3.11 (d, J=19.0 Hz, 1H), 3.04(td, J=14.5, 5.0 Hz, 1H), 2.73 (dd, J=12.3, 4.8 Hz, 2H), 2.63 (dd,J=18.9, 5.9 Hz, 1H), 2.46 (td, J=12.7, 5.3 Hz, 1H), 2.41 (dd, J=6.5, 1.8Hz, 2H), 2.33 (dt, J=14.5, 3.2 Hz, 1H), 2.08 (td, J=12.3, 3.8 Hz, 1H),1.92 (ddd, J=13.3, 5.0, 2.8 Hz, 1H), 1.57 (ddd, J=26.4, 13.3, 3.5 Hz,2H), 0.86 (p, J=5.7 Hz, 1H), 0.60-0.53 (m, 2H), 0.15 (q, J=4.8 Hz, 2H);¹³C NMR (151 MHz, CDCl₃) δ 206.7, 147.6, 133.9, 131.9, 131.5, 122.6,120.2, 91.7, 69.9, 61.7, 59.2, 51.1, 43.3, 36.1, 31.4, 30.8, 23.2, 9.4,4.1, 3.9; ¹⁹F NMR (376 MHz, CDCl₃) δ 38.3; ESI-MS (m/z): 424 [MH]⁺.

(S)-2-Azido-3-(4-((fluorosulfonyl)oxy)phenyl)propanoic acid (12-32). Theabove procedure was performed starting with the dicyclohexyl amine saltof (S)-2-azido-3-(4-((fluorosulfonyl)oxy)phenyl)propanoic acid and 2equiv Et₃N. The product was obtained as a yellow oil (0.65 g, 89%yield). ¹H NMR (400 MHz, CDCl₃) δ 8.02 (bs, 1H), 7.38 (m, 2H), 7.31 (m,2H), 4.20 (q, J=4.4 Hz, 1H), 3.23 (dd, J=14.4 Hz, 3.2 Hz, 1H), 3.05 (dd,J=14.4 Hz, 8.8 Hz, 1H); ¹³C NMR (101 MHz, CDCl₃) δ 174.4, 149.2, 136.7,131.3, 121.1, 62.6, 36.5; ¹⁹F NMR (376 MHz, CDCl₃) δ +37.2; ESI-MS(m/z): 262 [(M-N₂)H]⁺, 312[(M+Na)]⁺.

(E)-2-((4-((Fluorosulfonyl)oxy)phenyl)diazenyl)benzoic acid (12-33). Theabove procedure was performed with 2 equiv Et₃N. The crude product waspurified by quick filtration through silica gel, washing away impuritieswith 50% EtOAc in hexanes and eluting the product with 70% EtOAc inhexanes. R_(f) (1:1 hexane:EtOAc) 0.19. The desired product was obtainedas an orange solid in (7.16 g, 88% yield). mp 101-102° C. ¹H NMR (400MHz, CDCl₃) δ 8.46-8.36 (m, 1H), 8.04-7.92 (m, 3H), 7.76-7.68 (m, 2H),7.58 (d, J=8.5 Hz, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 166.5, 152.4, 151.0,149.7, 134.0, 133.4, 133.2, 127.7, 125.5, 122.6, 116.3; ¹⁹F NMR (376MHz, CDCl₃) δ 38.5; ESI-MS (m/z): 262 [M−H].

4-(5-Thioxo-4,5-dihydro-1H-tetrazol-1-yl)phenyl fluorosulfate (12-34).The above procedure was performed with 1.2 equiv Et₃N. The product wasobtained as a pink solid (6.29 g, 98% yield). mp 130-131° C. (dec.). ¹HNMR (400 MHz, DMSO-d₆) δ 8.18-8.12 (m, 2H), 7.92-7.86 (m, 2H); ¹³C NMR(101 MHz, CDCl₃) δ149.3, 134.2, 126.9, 126.5, 122.4; ¹⁹F NMR (376 MHz,CDCl₃) δ 39.8; ESI-MS (m/z): 277 [MH]⁺.

2-Oxo-7-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-2H-chromen-6-ylfluorosulfate (12-35). The above procedure was performed starting withesculin hydrate (540 mg, 1.5 mmol) and approximately 10.6 equiv Et₃N(added in 2 portions) in CH₂Cl₂:water (9 mL:6 mL, overall 0.1 M insubstrate). The crude product was purified by column chromatography onsilica gel [99:1-97:3 CH₃CN:water, R_(f) (95:5 CH₃CN:water) 0.49]. Afterevaporation from acetonitrile solution, the desired product was obtainedas a white solid (469 mg, 74% yield). mp 145-147° C. ¹⁹F NMR indicatedthe presence of a mixture of two conformational isomers in a 9:1 ratio.¹⁹F NMR (376 MHz, DMSO-d₆) δ 41.4 (major), 38.5 (minor); ¹H NMR (majorisomer) (500 MHz, DMSO-d₆) δ 7.55 (d, J=9.7 Hz, 1H), 7.45 (s, 1H), 7.30(s, 1H), 6.16 (d, J=9.7 Hz, 1H), 4.91 (d, J=5.0 Hz, 1H), 4.67 (d, J=4.5Hz, 1H), 4.63 (d, J=5.4 Hz, 1H), 4.61 (d, J=7.3 Hz, 1H), 4.12 (t, J=5.8Hz, 1H), 3.27 (ddd, J=11.7, 5.5, 2.1 Hz, 1H), 3.05 (dt, J=11.6, 5.8 Hz,1H), 2.99-2.94 (m, 1H), 2.86-2.82 (m, 2H), 2.79-2.73 (m, 1H); ¹³C NMR(major isomer) (126 MHz, DMSO-d₆) δ 159.3, 147.7, 144.9, 143.2, 139.8,119.6, 117.9, 115.8, 111.6, 100.9, 77.3, 76.8, 73.1, 69.5, 60.5; ESI-MS(m/z): 423 [MH]⁺.

III. Representative Procedure Using NaH as Base. Mesityl Fluorosulfonate(12-26)

The starting mesityl phenol (2.89 g, 21 mmol) was dissolved in dry THEand NaH (1.26 g of 60% suspension in mineral oil, 31.5 mmol, 1.5 equiv)was added under inert atmosphere at 0° C. The reaction mixture waswarmed to room temperature and stirred for 15-30 min. After evolution ofH₂ ceased, the mixture was cooled to 0° C., the inert gas inlet wasremoved, a low vacuum was created, and SO₂F₂ gas was introduced from aballoon with vigorous stirring. After 15 min at 0° C., the reactionmixture was warmed to room temperature. After 1 hour, GC-MS showed 97%conversion to the fluorosulfate, a residual amount of starting phenol,and a small amount of disulfate (less than 1%). The reaction mixture wasopened to air, quenched with 1N HCl (about 35 mL) until a pH of 3-4 wasestablished, extracted with EtOAc (2×50 mL), washed with brine, driedover MgSO₄ and concentrated. The crude product was further purified byshort column chromatography to give a colorless oil (4.37 g, 94% yield).R_(f) (95:5 hexane:EtOAc) 0.72.

Fluorosulfate of (+)-α-tocopherol (12-36). (+)-α-Tocopherol wasextracted from a commercially available mixture of (+)-α-tocopherol andvegetable oil according to the literature procedure of Isso, B.; Ryan,D. Eur. J. Lipid Sci. Technol. 2012, 114, 927-932. The vitamin E/oilmixture (20 g, approximately 60% vitamin E) was dissolved in hexane (40mL). A mixture of 20 mL acetonitrile and 20 mL methanol was added andthe resulting mixture was vortexed for 1 min, then allowed to stand for5 min to separate. The top acetonitrile-methanol layer was isolated,washed with hexanes, and concentrated, providing 13.4 g of(+)-α-tocopherol (77% recovery). R_(f) (95:5 hexane:EtOAc) 0.56.

After subjection to the above SO₂F₂ reaction procedure, the crudefluorosulfate of (+)-α-tocopherol was purified by column chromatography(100:0 to 95:5 hexane:Et₂O). R_(f)(95:5 hexane:EtOAc) 0.90. The productwas obtained as a thick colourless oil (9.82 g, 82% yield). ¹H NMR (400MHz, CDCl₃) δ 2.62 (t, J=6.8 Hz, 2H), 2.25 (s, 3H), 2.22 (s, 3H), 2.13(s, 3H), 1.83 (ddt, J=35.5, 13.2, 6.8 Hz, 2H), 1.66-1.50 (m, 3H),1.51-1.36 (m, 4H), 1.36-1.22 (m, 8H), 1.27 (s, 3H), 1.20-1.05 (m, 6H),0.93-0.85 (m, 12H); ¹³C NMR (101 MHz, CDCl₃) δ 151.1, 142.0, 127.6,126.2, 124.5, 118.5, 75.9, 40.1, 39.5, 37.6, 37.5, 37.4, 33.0, 32.8,31.0, 28.1, 25.0, 24.6, 24.0, 22.9, 22.8, 21.1, 20.8, 19.9, 19.8, 13.7,12.9, 12.1; ¹⁹F NMR (376 MHz, CDCl₃) δ 40.8; Anal. Calcd. forC₂₉H₄₉FO₄S: C, 67.93; H, 9.63; F, 3.71. Found: C, 67.73; H, 9.56; F,3.72.

2,5-Dimethyl-4-(morpholinomethyl)phenyl fluorosulfate (12-37). In theabove procedure, 3 equiv NaH was used with2,5-dimethyl-4-(morpholinomethyl)phenol-HCl hydrate as the startingmaterial. Column chromatography (9:1 to 7:3 hexane:EtOAc) gave thedesired product as a white solid (2.4 g, 54% yield). mp 93.5-95.5° C. ¹HNMR (400 MHz, CDCl₃) δ 7.20 (s, 1H), 7.08 (s, 1H), 3.69 (br s, 4H), 3.41(s, 2H), 2.43 (br s, 4H), 2.35 (s, 3H), 2.32 (s, 3H); ¹³C NMR (101 MHz,CDCl₃) δ 147.9, 137.7, 136.8, 133.4, 127.1, 122.4, 67.2, 60.6, 53.8,19.0, 15.7; ¹⁹F NMR (376 MHz, CDCl₃) δ 38.4; ESI-MS (m/z): 304 [MH]⁺.

IV. Representative Procedure Using DBU as Base

Dinaphtho[2,1-d:1′,2′-f][1,3,2]dioxathiepine 4,4-dioxide (12-38).Binaphthol (1.0 g, 3.5 mmol) was dissolved in acetonitrile (35 mL, 0.1 Min substrate). The reaction flask was sealed with a septum, air wasevacuated, and a balloon filled with SO₂F₂ gas was introduced, followedby addition of DBU by syringe (1.1 mL, 7.2 mmol, 2.05 equiv). Thereaction mixture was vigorously stirred under SO₂F₂ atmosphere at roomtemperature for several hours, monitoring by GC-MS and TLC. Uponcompletion, the reaction was diluted with EtOAc (50 mL), washed with 1NHCl (2×25 mL), brine 10 mL), dried over MgSO₄, and concentrated. Thedesired product was obtained as a white solid (1.21 g, quant.). mp198-199° C. R_(f) (7:3 hexane:EtOAc) 0.33. GC-MS (t_(R)): 9.6 min; EI-MS(m/z): 348 [M]⁺. NMR spectra matched the data reported previously byKoy, C.; Michalik, M.; Oehme, G.; Alm, J.; Kempe, R. Sulfur Letters,1998, 21(2), 75-88.

Ex. 1(K). Fluorosulfates and Sulfates Via Silyl-Fluoride Metathesis

Section A. Representative procedure for the synthesis of fluorosulfatesfrom silylated phenols and sulfuryl fluoride gas in the presence of DBU.4-(methylamino)phenyl fluorosulfonate (13A-1). TBS protected phenol (2.4g, 10.15 mmol) was dissolved in dry acetonitrile (20 mL, 0.5 M insubstrate). The reaction flask was sealed with septa, air was evacuated,a balloon filled with SO₂F₂ gas was introduced, and DBU was injected bysyringe (145 μL, 1 mmol). The reaction mixture was vigorously stirred atroom temperature for several hours, monitoring by GC-MS or LC-MS. Uponcompletion, the reaction was quenched with 1N HCl (50 mL) and extractedwith EtOAc (2×100 mL). The organic extracts were washed with brine (35mL), dried over MgSO₄, and concentrated. The product was obtained as abrown oil, which solidified upon standing (2.05 g, 99% yield). mp 37-40°C. R_(f) (7:3 hexane:EtOAc): 0.55. ¹H NMR (400 MHz, CDCl₃) δ 7.15-7.12(m, 2H), 6.59 (m, 2H), 4.12 (bs, 1H), 2.83 (s, 3H); ¹³C NMR (101 MHz,CDCl₃) δ 148.9, 141.4, 121.6, 112.8, 30.7; ¹⁹F NMR (376 MHz, CDCl₃) δ+35.3; GC (t_(R)): 5.2 min; EI-MS (m/z): 205 [M]⁺.

Polar aprotic solvents (acetonitrile, NMP and DMF) facilitate thetransformation. Aprotic solvents (CH₂Cl₂, chloroform, trifluorotoluene,THF) can be used as well, at the cost of longer reaction times. Thechoice of the phenolic silyl ether group also has a pronounced effect onreaction rate. Reactions involving TMS-protected phenols usually require1-4 h to reach completion, whereas reactions with the bulkier TBS grouprequire 6-8 h. The DBU catalyst can be replaced with DBN or BEMP.

Benzene-1,3,5-triyl trifluorosulfonate (13A-2). The use of1,3,5-tris((tert-butyldimethylsilyl)oxy)benzene (1.0 g, 2.1 mmol) inCH₃CN:THF (10 mL+10 mL) and 30 mol % of DBU gave a white solid.Additional purification was performed by column chromatography (9:1hexane:EtOAc) to give the desired compound as a white solid (0.69 g, 86%yield). mp 96-98° C. R_(f) (9:1 hexane:EtOAc) 0.62. ¹H NMR (400 MHz,CDCl₃) δ 7.53 (s, 3H); ¹³C NMR (101 MHz, CDCl₃) δ 150.3, 115.7; ¹⁹F NMR(376 MHz, CDCl₃) δ 39.5; GC-MS (t_(R)): 4.9 min; EI-MS (m/z): 372 [M]⁺.

2,2-Dioxidobenzo[d][1,3,2]dioxathiol-4-yl fluorosulfonate (13A-3).1,2,3-Tris((trimethylsilyl)oxy)benzene (3.42 g, 10 mmol) in CH₃CN (100mL) was treated with DBU (30 mol %, 450 μL) and SO₂F₂ gas was introducedto the solution cooled to 0° C. in an ice bath. The reaction mixture wasvigorously stirred for 4-5 hours, after which EtOAc was used instead ofether for the reaction workup. The crude product was purified by columnchromatography (95:5 to 7:3 hexane:EtOAc) to give the desired compoundas a colorless oil (1.9 g, 70% yield). R_(f) (7:3 hexane:EtOAc) 0.66. ¹HNMR (400 MHz, CDCl₃) δ 7.39-7.29 (m, 1H); ¹³C NMR (101 MHz, CDCl₃) δ143.72, 134.49, 132.87, 125.81, 119.03, 112.24; ¹⁹F NMR (376 MHz, CDCl₃)δ 40.2; GC-MS (t_(R)): 5 min; EI-MS (m/z): 270 [M]⁺.

(S)-tert-Butyl(3-(2,2-dioxidobenzo[d][1,3,2]dioxathiol-5-yl)-1-(methoxy(methyl)amino)-1-oxopropan-2-yl)carbamate(13A-4). The bis-TBS protected phenol (1.62 g, 2.85 mmol) was taken upin CH₃CN (20 mL). DBU (20 mol %; 85 μL) and SO₂F₂ gas were introduced atroom temperature as described above, and the reaction mixture wasvigorously stirred for 18 h. The crude material was passed through ashort plug of silica gel, eluting with hexane:EtOAc (3:1 to 1:1) to givethe desired product as a pure colorless oil that solidified on standing(1.1 g, 96% yield). mp 108-111° C. R_(f) (3:2 hexane:EtOAc) 0.41. ¹H NMR(500 MHz, CDCl₃) δ 7.04 (d, J=8.4 Hz, 1H), 7.02 (s, 1H), 6.96 (d, J=8.5Hz, 1H), 5.49 (d, J=8.4 Hz, 1H), 4.88-4.80 (m, 1H), 3.68 (s, 3H), 3.12(s, 3H), 3.01 (dd, J=13.7, 5.4 Hz, 1H), 2.80 (dd, J=13.8, 7.8 Hz, 1H),1.30 (s, 9H); ¹³C NMR (126 MHz, CDCl₃) δ 171.4, 155.1, 142.3, 141.3,135.1, 126.2, 112.7, 111.3, 79.7, 61.6, 51.4, 38.4, 32.0, 28.1; EI-MS(m/z): 425 [M+Na]⁺.

Section B. Representative procedure for the synthesis of sulfates fromsilylated phenols and fluorosulfates in the presence of DBU. Silylatedether (1 equiv) was dissolved in acetonitrile (0.1-0.5 M in substrate),fluorosulfate (1 equiv) was added, and the reaction mixture was stirredfor several minutes to obtain a homogeneous solution. DBU (10-30 mol %)was then added and the reaction mixture was stirred at room temperature(unless noted otherwise) for several hours, monitoring reaction progressby LC-MS or TLC. Note that the reaction vessel must be vented to allowfor escape of the volatile silyl fluoride byproduct. Upon completion,the reaction was quenched with 1N HCl (unless amine functionality waspresent in the substrate), extracted with EtOAc, washed with brine,dried over MgSO₄, and concentrated. Crude products were purified bycolumn chromatography when necessary.

Polar aprotic solvent (acetonitrile, NMP, DMF) facilitate thetransformation. When the silyl ether has poor solubility inacetonitrile, THE can be used as a co-solvent. The choice of the silylgroup on the phenol component should be adjusted depending on the stericproperties of the fluorosulfate component. Since sterically hinderedfluorosulfates require prolonged reaction time and elevatedtemperatures, the use of more stable TBS-protected phenols as couplingpartners in this case generally provide cleaner transformations. Morereactive fluorosulfates give cleaner transformations with more reactiveTMS-protected phenols. In cases when reactive fluorosulfates are coupledwith slow TBS protected phenols, the symmetric product of homocouplingof fluorosulfates can be observed in various amounts. DBU catalyst canbe replaced with DBN or BEMP.

(8R,9S,13S,14S,17R)-17-Ethynyl-17-hydroxy-13-methyl-7,8,9,11,12,13,14,15,16,17-decahydro-6H-cyclopenta[a]phenanthren-3-yl((R)-2,5,7,8-tetramethyl-2-((4R,8R)-4,8,12-trimethyltridecyl)chroman-6-yl)sulfate (13B-1). Prepared from fluorosulfate (493 mg, 0.96 mmol),acetonitrile (4 mL), the TBS-protected steroid (381 μL, 0.96 mmol), andDBU (30 L, 0.19 mmol), with stirring at 80° C. overnight. Columnchromatography (9:1 to 7:3 hexane:EtOAc) provided the desired product asa white foam (0.73 g, 98% yield). R_(f) (7:3 hexane:EtOAc) 0.47. ¹H NMR(600 MHz, CDCl₃) δ 7.33 (d, J=8.1 Hz, 1H), 7.11 (d, J=7.3 Hz, 1H), 7.05(s, 1H), 2.88 (br s, 1H), 2.62 (br s, 2H), 2.43-2.32 (m, 2H), 2.27 (s,3H), 2.23 (s, 3H), 2.12 (s, 3H), 2.10-2.00 (m, 2H), 2.00-1.88 (m, 2H),1.88-1.68 (m, 4H), 1.66-1.28 (m, 20H), 1.27 (s, 6H), 1.20-1.03 (m, 6H),0.95-0.81 (m, 12H).

2,5-Dimethyl-4-(morpholinomethyl)phenyl phenyl sulfate (13B-2). Preparedfrom fluorosulfate (883 mg, 2.91 mmol), acetonitrile (6 mL, 0.5 M insubstrate), trimethyl(phenoxy)silane (527 μL, 2.91 mmol), and DBU (44μL; 0.28 mmol), with stirring at room temperature for 5 h and quenchingof the reaction with sat. NaHCO₃ (5 mL). Column chromatography (9:1 to7:3 hexane:EtOAc) gave the desired product as a colorless oil (0.95 g,86% yield). R_(f) (7:3 hexane:EtOAc) 0.42. ¹H NMR (400 MHz, CDCl₃) δ7.40-7.25 (m, 5H), 7.12 (s, 1H), 7.06 (s, 1H), 3.63 (t, J=4.6 Hz, 4H),3.35 (s, 2H), 2.37 (t, J=4.5 Hz, 4H), 2.28 (s, 3H), 2.20 (s, 3H); ¹³CNMR (101 MHz, CDCl₃) δ 150.3, 147.8, 137.0, 135.5, 133.0, 129.9, 127.5,127.2, 122.3, 121.2, 66.9, 60.4, 53.5, 18.8, 15.6; ESI-MS (m/z): 378[MH]⁺.

Bis(2-oxo-2H-chromen-7-yl) (propane-2,2-diylbis(4,1-phenylene))bis(sulfate) (15B-3). Prepared from fluorosulfate (513 mg, 2.1 mmol),acetonitrile (5 mL), TMS-protected bisphenol A (373 mg, 1 mmol), and DBU(30 μL; 0.2 mmol), with stirring at room temperature for 4 h. Theproduct was isolated directly as a precipitated white solid (filteredand washed with acetonitrile, 0.5 g, 75% yield). mp 169-174° C. ¹H NMR(500 MHz, CDCl₃) δ 7.67 (d, J=9.6 Hz, 2H), 7.51 (d, J=8.4 Hz, 2H),7.26-7.20 (m, 12H), 6.41 (d, J=9.6 Hz, 2H), 1.66 (s, 6H); ¹³C NMR (126MHz, CDCl₃) δ 159.7, 154.8, 152.2, 145.0, 148.5, 142.5, 129.4, 128.7,120.8, 118.2, 117.6, 117.4, 110.1, 43.0, 30.9; ESI-MS (m/z): 677 [MH]⁺.

(S)-Benzyl2-amino-3-(4-((((6-methylpyridin-3-yl)oxy)sulfonyl)oxy)phenyl)propanoate(13B-4). Prepared from fluorosulfate (478 mg, 2.5 mmol), acetonitrile(10 mL), TBS-protected tyrosine derivative (964 mg, 2.5 mmol), and DBU(75 μL, 0.5 mmol), with stirring at room temperature for 4 h. Columnchromatography (9:1 CH₂Cl₂/MeOH) gave the desired compound as a yellowoil (0.9 g, 83% yield). R_(f) (9:1 CH₂Cl₂:MeOH) 0.44. ¹H NMR (600 MHz,CDCl₃) δ 8.43 (d, J=2.9 Hz, 1H), 7.52 (dd, J=8.5, 3.0 Hz, 1H), 7.36-7.27(m, 6H), 7.20 (d, J=8.3 Hz, 1H), 7.18 (m, 3H), 5.12 (q, J=12.2 Hz, 2H),3.74 (t, J=6.5 Hz, 1H), 3.05 (dd, J=13.6, 5.7 Hz, 1H), 2.90 (dd, J=13.6,7.3 Hz, 1H), 2.57 (s, 3H), 1.66 (br s, 2H); ¹³C NMR (101 MHz, CDCl₃) δ174.6, 158.1, 149.1, 145.5, 142.0, 137.2, 135.4, 131.1, 129.0, 128.7,128.6, 128.6, 124.1, 120.9, 66.9, 55.6, 40.2, 24.0; EI-MS (m/z): 443[MH]⁺.

4-(2-Amino-2-oxoethyl)phenyl(4-(1-((tert-butyldimethylsilyl)oxy)-2-(methylamino)ethyl)-phenyl)sulfate (13B-5). Prepared from fluorosulfate (483 mg, 2.07 mmol),acetonitrile (10 mL), TBS-protected phenol (818 mg, 2.07 mmol), and DBU(75 μL, 0.5 mmol), with stirring at room temperature overnight. Columnchromatography (reversed-phase, 0-60% gradient of acetonitrile in water)gave the desired product as a white solid (0.8 g, 75% yield). mp108-110° C. ¹H NMR (600 MHz, CDCl₃) δ 7.39 (d, J=8.6 Hz, 2H), 7.34 (d,J=8.6 Hz, 2H), 7.29 (d, J=8.8 Hz, 2H), 7.28 (d, J=8.7 Hz, 2H), 5.56 (d,J=122.4 Hz, 2H), 4.85 (dd, J=8.3, 4.0 Hz, 1H), 3.59 (s, 2H), 2.75 (dd,J=12.1, 8.2 Hz, 1H), 2.64 (dd, J=12.1, 3.9 Hz, 1H), 2.45 (s, 3H), 0.89(s, 9H), 0.05 (s, 3H), −0.14 (s, 3H); ¹³C NMR (151 MHz, CDCl₃) δ 172.5,149.7, 149.5, 143.4, 134.5, 131.2, 127.8, 121.7, 120.9, 73.4, 60.7,42.5, 36.4, 25.9, −4.4, −4.8; ESI-MS (m/z): 495 [MH]⁺.

1,4-Phenylene bis(4-allyl-2-methoxyphenyl) bis(sulfate) (13-B6).Prepared from fluorosulfate (1.23 g, 5 mmol), acetonitrile (5 mL),1,4-bis((trimethylsilyl)oxy)benzene (636 mg, 2.5 mmol), and DBU (75 μL,0.5 mmol), with stirring at 50° C. for 5 h. The crude product waspurified on a short column chromatography (9:1 to 8:2 hexane:EtOAc) togive a white solid (1.3 g, 92% yield). mp 62-65° C. R_(f) (9:1hexane:EtOAc) 0.22. ¹H NMR (400 MHz, CDCl₃) δ 7.48 (s, 4H), 7.20 (d,J=8.2 Hz, 2H), 6.81 (s, 2H), 6.78 (d, J=8.3 Hz, 2H), 5.93 (ddt, J=18.1,9.5, 6.7 Hz, 2H), 5.13 (s, 2H), 5.09 (d, J=4.3 Hz, 2H), 3.78 (s, 6H),3.38 (d, J=6.7 Hz, 4H). ¹³C NMR (101 MHz, CDCl₃) δ 151.2, 149.1, 141.4,137.7, 136.6, 122.9, 122.5, 120.9, 116.8, 113.5, 56.0, 40.1; EI-MS(m/z): 563 [MH]⁺.

5-Chloroquinolin-8-yl (4-(methylamino)phenyl) sulfate (13-B7). Preparedfrom fluorosulfate (786 mg, 3 mmol), acetonitrile (12 mL), TBS-protectedphenol (711 mg, 3 mmol), and DBU (90 μL, 0.6 mmol), with stirring atroom temperature overnight. Column chromatography (9:1 to 6:4hexane:EtOAc) gave the product as a yellow solid (0.7 g, 61% yield). mp99-100.5° C. R_(f) (1:1 hexane:EtOAc) 0.62. ¹H NMR (400 MHz, CDCl₃) δ9.08 (dd, J=4.3, 1.7 Hz, 1H), 8.58 (dd, J=8.6, 1.7 Hz, 1H), 7.64-7.53(m, 3H), 7.31 (d, J=8.9 Hz, 2H), 6.58 (d, J=8.9 Hz, 2H), 2.83 (s, 3H);¹³C NMR (151 MHz, CDCl₃) δ 151.9, 148.7, 145.6, 141.7, 141.5, 133.1,130.4, 127.6, 125.9, 123.1, 122.4, 121.1, 112.5, 30.8; ESI-MS (m/z): 365[MH]⁺.

Ex. 1(L). Synthesis of Enol Fluorosulfates

3,4-Dihydronaphthalen-1-yl sulfonate from silylated enol ether (15-1). Around-bottom flask was charged with((3,4-dihydronaphthalen-1-yl)oxy)trimethylsilane (436 mg, 2 mmol) anddry CH₂Cl₂ (5 mL), then sealed with septa. Air was evacuated and SO₂F₂gas was introduced in a balloon, followed by addition of 1M solution ofBEMP in hexane by syringe (200 μL, 0.2 mmol, 10 mol %). The reactionmixture was stirred at room temperature overnight, monitoring by TLC.Upon completion, the solvent was removed by rotary evaporation and thecrude product was purified by column chromatography (10:1 hexane:EtOAc)to give a colorless oil (350 mg, 77% yield). R_(f) (8:2 hexane:EtOAc)0.67. ¹H NMR (400 MHz, CDCl₃) δ 7.40-7.37 (m, 1H), 7.31-7.26 (m, 2H),7.22-7.19 (m, 1H), 6.11 (dt, J=8 Hz, 4 Hz, 1H), 2.90 (t, J=8 Hz, 2H),2.57-2.51 (m, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 147.2, 136.4, 129.4,128.0, 127.0, 121.2, 117.6, 26.9, 22.2; ¹⁹F NMR (376 MHz, CDCl₃) δ+39.1; GC-MS (t_(R)): 5.2 min; EI-MS (m/z): 228 [M]⁺.

Note: The analogous reaction in acetonitrile was complete within severalminutes.

3,4-Dihydronaphthalen-1-yl sulfonate from lithium enolate generated insitu (15-1). A Schlenk flask was charged with α-tetralone (333 μL, 2.5mmol) in dry THE (5 mL) and cooled to −78° C. under dry atmosphere.LHMDS (3.75 mL of 1M in THF, 3.75 mmol) was added by syringe and thereaction mixture was stirred at −78° C. for an additional 30 min. Lowvacuum was then applied, SO₂F₂ was introduced from a balloon, thereaction was allowed to warm to 0° C., and stirring continued for 1 hourat that temperature. The reaction was monitored by TLC and GC-MS. Uponcompletion, the reaction was quenched with water (5 mL), extracted withEtOAc (2×10 mL), washed with brine, and concentrated. Purification bychromatography on a short silica column (10:1 hexane:EtOAc) gave thedesired compound as a colorless oil (400 mg, 75% yield).

Ex. 1(M). Synthesis of Sulfamoyl Fluorides

General Procedure. Secondary amine (1 equiv), DMAP (0.5-1 equiv) andtriethylamine (2 equiv) were mixed in CH₂Cl₂ (0.33 M) in a round-bottomflask filled to one-third capacity. The flask was sealed with septa, airwas evacuated, and SO₂F₂ gas was introduced from a balloon. The reactionmixture was stirred vigorously at room temperature for 3-18 h, andreaction progress was monitored by GC or LC-MS. Upon completion, themixture was concentrated, dissolved in EtOAc, washed with 1N HCl andbrine, dried over MgSO₄, and concentrated to give the desired compound,usually in quite pure form. In some cases, additional purification wasperformed by passage through a short silica gel column.

Activated cyclic amines react rapidly with SO₂F₂; in some cases, coolingof the reaction mixture with a water bath is required. Acyclic aminesrequire 1-1.5 equiv DMAP. When 1.5 equiv of DMAP are used, the additionof triethylamine or other extra base is not required. Activated aminescan even react in buffer at pH 8 (phosphate, borate, or HEPES buffers).

Di(prop-2-yn-1-yl)sulfamoyl fluoride (17-1) was prepared with 1 equivDMAP and 2 equiv Et₃N. Brown impurities in the product were removed bypassing the material through a short plug of silica gel (9:1hexane:EtOAc). The product was obtained as a pink oil (3.3 g, 76% yield,accounting for CH₂Cl₂ impurity by ¹H NMR). ¹H NMR (400 MHz, CDCl₃) δ4.28 (t, J=2.2 Hz, 4H), 2.47 (t, J=2.4 Hz, 2H); ¹³C NMR (101 MHz, CDCl₃)δ 75.7, 74.8 (d, J=1.6 Hz), 37.9 (d, J=1.3 Hz; ¹⁹F NMR (376 MHz, CDCl₃)δ 46.4; R_(f) (9:1 hexane:EtOAc) 0.29; EI-MS (m/z): 174 [M]⁺.

Diallylsulfamoyl fluoride (17-2) was prepared with 1 equiv DMAP and 2equiv Et₃N, obtained as a yellow oil (2.9 g, 65% yield). ¹H NMR (400MHz, CDCl₃) δ 5.81 (ddtd, J=16.8, 10.2, 6.5, 1.1 Hz, 2H), 5.36-5.28 (m,4H), 3.94 (d, J=6.3 Hz, 4H); ¹³C NMR (101 MHz, CDCl₃) δ 130.7 (d, J=1.6Hz), 120.9, 50.8 (d, J=2.0 Hz); ¹⁹F NMR (376 MHz, CDCl₃) δ 50.00; R_(f)(7:3 hexane:EtOAc): 0.72; GC-MS (t_(R)): 3.7 min; EI-MS (m/z): 178 [M]⁺.

Bis(2-azidoethyl)sulfamoyl fluoride (17-3) was prepared from thebis(2-azidoethyl)amine mesylate salt, 1 equiv DMAP, and 3 equiv Et₃N,isolated as a yellow oil (4.49 g, 76% yield). ¹H NMR (400 MHz, CDCl₃) δ3.64-3.59 (m, 4H), 3.59-3.54 (m, 4H); ¹³C NMR (101 MHz, CDCl₃) δ 50.2(d, J=1.9 Hz), 49.7 (d, J=1.6 Hz); ¹⁹F NMR (376 MHz, CDCl₃) δ 50.0;R_(f) (9:1 hexane:EtOAc): 0.52; HRMS (ESI-TOF) (m z): [M+H]⁺ calcd forC₄H₈FN₇O₂S, 238.0517; found 238.0524.

(2,2-Dimethoxyethyl)(methyl)sulfamoyl fluoride (17-4) was prepared with1 equiv DMAP and 2 equiv Et₃N, obtained as a colorless oil (3.4 g, 88%yield). ¹H NMR (400 MHz, CDCl₃) δ 4.50 (t, J=5.4 Hz, 1H), 3.41 (s, 6H),3.35 (dd, J=5.4, 2.0 Hz, 2H), 3.08 (d, J=2.3 Hz, 3H); ¹³C NMR (101 MHz,CDCl₃) δ 103.0 (d, J=2.4 Hz), 55.0, 52.8 (d, J=2.1 Hz), 37.9 (d, J=1.5Hz); ¹⁹F NMR (376 MHz, CDCl₃) δ 50.0; R_(f) (7:3 hexane:EtOAc): 0.44;EI-MS (m/z): 201 [M]⁺.

4-(2-Azidoacetyl)piperazine-1-sulfamoyl fluoride (17-5) was preparedfrom 2-azido-1-(piperazin-1-yl)ethanone-TFA salt, 0.5 equiv DMAP, and 5equiv Et₃N. The product was obtained as a pink solid (4.4 g, 70% yield).mp 92-94° C. ¹H NMR (400 MHz, CDCl₃) δ 3.93 (br s, 2H), 3.72 (br s, 2H),3.55-3.30 (m, 6H); ¹³C NMR (101 MHz, CDCl₃) δ 165.9, 50.7, 46.7, 44.0,40.8; ¹⁹F NMR (376 MHz, CDCl₃) δ 40.1; R_(f) (1:1 hexane:EtOAc): 0.44;GC-MS (t_(R)): 6.3 min; EI-MS (m/z): 251 [M]⁺.

6,7-Dimethoxy-1-phenyl-3,4-dihydroisoquinoline-2(1H)-sulfamoyl fluoride(17-6) was prepared with 1 equiv DMAP and 2 equiv Et₃N, obtained as awhite solid (0.6 g, 80% yield). mp 109-111° C. ¹H NMR (400 MHz, CDCl₃) δ7.35-7.32 (m, 3H), 7.26-7.19 (m, 2H), 6.70 (s, 1H), 6.44 (s, 1H), 6.11(s, 1H), 3.98-3.89 (m, 1H), 3.91 (s, 3H), 3.75 (s, 3H), 3.43-3.29 (m,1H), 3.21-3.12 (m, 1H), 2.75 (ddd, J=16.6, 4.4, 1.6 Hz, 1H); ¹³C NMR(101 MHz, CDCl₃) δ 148.8, 148.0, 139.8 (d, J=2.3 Hz), 129.1, 128.7,128.7, 125.4, 124.1, 111.4, 110.7, 60.5 (d, J=1.4 Hz), 56.1 (d, J=5.5Hz), 40.6 (d, J=2.7 Hz), 26.8; ¹⁹F NMR (376 MHz, CDCl₃) δ 50.0; R_(f)(7:3 hexane:EtOAc): 0.56; GC-MS (t_(R)): 8.3 min; EI-MS (m/z): 351 [M]⁺.

4-(Dibenzo[b,f][1,4]oxazepin-11-yl)piperazine-1-sulfamoyl fluoride(17-7) was prepared with 0.5 equiv DMAP and 2 equiv Et₃N, obtained as awhite solid (0.5 g, 73% yield). mp 142-144° C. ¹H NMR (400 MHz, CDCl₃) δ7.44 (d, J=6.2 Hz, 1H), 7.31 (s, 1H), 7.25-7.00 (m, 5H), 3.66 (s, 4H),3.58 (s, 4H); ¹³C NMR (101 MHz, CDCl₃) δ 159.6, 158.4, 151.8, 139.5,133.3, 130.8, 128.7, 127.3, 126.1, 125.6, 124.6, 123.1, 120.4, 46.6,46.65 (n); ¹⁹F NMR (376 MHz, CDCl₃) δ 38.6; R_(f) (9:1 hexane:EtOAc):0.48; EI-MS (m/z): 396 [MH]⁺.

4-((3-Methylbutylidene)amino)piperidine-1-sulfamoyl fluoride (17-8) wasprepared with 0.5 equiv DMAP and 2 equiv Et₃N. Upon completion, thereaction mixture was washed with water, dried, and concentrated. Theproduct was obtained as a yellow oil yield (1.8 g, 70% yield, accountingfor DMAP contamination in ¹H NMR). For characterization, the imine groupwas hydrolyzed (treatment with ^(i)PrOH/water mixture at 50° C. for 1.5hours) followed by treatment with MsOH to make the shelf stable salt(17-8a). EI-MS (m/z): 183 [MH]⁺.

The amine was then converted to the corresponding acetamide (17-8b)(treatment with Ac₂O/Py in CH₂Cl₂ at room temperature for 18 h, followedby acidification (1M HCl) and extraction with CH₂Cl₂). The amide wasobtained as a beige solid in 48% yield (3 steps from crude4-((3-methylbutylidene)amino)piperidine-1-sulfonyl fluoride, 160 mg). ¹HNMR (400 MHz, CDCl₃) δ 5.71 (s, 1H), 4.02-3.82 (m, 2H), 3.13 (t, J=12.6Hz, 2H), 2.04 (d, J=12.2 Hz, 2H), 1.97 (s, 3H), 1.63-1.46 (m, 2H); ¹³CNMR (101 MHz, CDCl₃) δ 169.8, 46.5, 45.6, 30.9, 23.5; ¹⁹F NMR (376 MHz,CDCl₃) δ (pm): 41.4; R_(f) (CH₂Cl₂/MeOH—9/1): 0.54; EI-MS (m/z): 225[MH]⁺.

4-(Hydroxydiphenylmethyl)piperidine-1-sulfamoyl fluoride (17-9). Theamine precursor (6.68 g, 25 mmol), DMAP (1.5 g, 12.5 mmol), and MgO (2.5g, 62.5 mmol) were mixed in a 4:1 CH₂Cl₂:water mixture (0.5 M insubstrate) in a 500 mL round-bottom flask. The flask was sealed withsepta, air was evacuated, and a SO₂F₂-filled balloon was introduced. Thereaction was vigorously stirred at room temperature for 6-18 h. Uponcompletion, the mixture was filtered through a short plug of CELITE©,and washed with water (50 mL) and then CH₂Cl₂ (200 mL). The organiclayer was washed with 1N HCl (50 mL), brine (50 mL), dried over MgSO₄,and concentrated. The product was obtained as a white solid (8.2 g, 94%yield). mp 109-112° C. ¹H NMR (400 MHz, CDCl₃) δ 7.61-7.42 (m, 4H),7.42-7.29 (m, 4H), 7.29-7.18 (m, 2H), 3.94 (d, J=9.4 Hz, 2H), 3.02 (brs, 2H), 2.58 (br s, 1H), 2.33 (s, 1H), 1.74-1.51 (m, 4H); ¹³C NMR (101MHz, CDCl₃) δ 145.0, 128.5, 127.1, 125.7, 79.4, 47.7, 43.3, 25.6; ¹⁹FNMR (376 MHz, CDCl₃) δ (pm): 39.2; R_(f) (7:3 hexane:EtOAc): 0.56; EI-MS(m/z): 332 [(M−H₂O)H]⁺.

1,4-Dioxa-8-azaspiro[4.5]decane-8-sulfamoyl fluoride (17-10) wasobtained as a yellow solid (5.6 g, 99% yield) using the same generalprocedure as described above for the synthesis of 17-9, but with1,4-dioxa-8-azaspiro[4.5]decane as the amine precursor. mp 87-89° C. ¹HNMR (400 MHz, CDCl₃) δ 3.97 (s, 2H), 3.62-3.53 (m, 2H), 1.81 (t, J=5.9Hz, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 105.5, 64.7, 45.8 (d, J=1.3 Hz),34.1 (d, J=1.2 Hz); ¹⁹F NMR (376 MHz, CDCl₃) δ 41.7; R_(f) (7:3hexane:EtOAc): 0.41; GC-MS (t_(R)): 5.35 min; EI-MS (m/z): 225 [M]⁺.

Example 2. Modified Antibiotics

ArOSO₂F is a non-polar functional group on an aromatic ring. It is anelectrophile that can coexist with nucleophiles and withstand biologicalsystems. ArOSO₂F is very stable and can selectively react with differentprotein targets. Its non-polar functionality means introduction of thefunctional group on the parent minimally impacts, if at all, theaffinity for the parent molecule.

Any known small molecule drugs which have one or more aromaticsubstitution can be readily converted to ArOSO₂F. Many antibioticsinclude functional groups such as aryl-OH, amino groups, and the like,which can be derivatized to introduce an SO₂F group (e.g., OSO₂F,NCH₂CH₂SO₂F, or NSO₂F) into the antibiotic structure. In this study,five fluorosulfonyl antibiotic derivatives (cephalosporin derivative10-2, ciprofloxacin derivative 10-7, and three vancomycinderivatives—vancomycin-SF, vancomycin-SF-1 and vancomycin-SF-2, thelatter two of which include N-propargyl groups as potential reactants incopper-catalyzed azide/alkyne click coupling reactions) were evaluatedfor antimicrobial activity against E. coli and/or B. subtilis incomparison to the non-modified versions of the antibiotics. See FIG. 20for the structures of the antibiotic compounds.

The antibiotic compounds were serially diluted in LB medium (10 g/Ltryptone, 5 g/L yeast extract, 10 g/L NaCl in deionized water) on a96-well culture plate from DMSO stock solution. Bacteria were inoculatedinto the wells and allowed to grow overnight at 37° C., 300 rpm.Overnight growth was assessed by measuring the turbidity of the media byabsorbance at 605 nm in a plate reader. The results for Bacillussubtilis are shown in Table 2, and the results for E. coli are shown inTable 3. Low turbidity (low optical density at 605 nm, OD₆₀₅) relativeto medium or vehicle alone indicates antibacterial activity.

TABLE 2 Bacillus subtilis (Gram positive) results. OD₆₀₅ afterAntibiotic Structure overnight 50 μM in FIG. 20 growth LB medium onlyN/A 0.425 Vehicle (DMSO) N/A 0.422 Cephalosporin A 0.527Cephalosporin-SF B 0.16 Ciprofloxacin C 0.036 Ciprofloxacin-SF D 0.036Vancomycin E 0.036 Vancomycin-FS F 0.037 Vancomycin-FS-1 G 0.037Vancomycin-FS-2 H 0.049

TABLE 3 Escherichia coli (Gram negative) results. OD₆₀₅ after AntibioticStructure overnight 20 μM in FIG. 20 growth Vehicle (DMSO) — 0.325Cephalosporin A 0.253 Cephalosporin-SF B 0.339 Ciprofloxacin C 0.036Ciprofloxacin-SF D 0.037

The results in Tables 2 and 3 clearly indicate that thefluorosulfonylated antibiotics exhibit similar activity to thenon-derivatized antibiotics, although the cephalosporin derivativeappears to have improved activity against B. subtilis relative tocephalosporin itself in these tests.

Example 3. Modified Tyrosine and Peptides Prepared Therewith

Fluorosulfate-Fmoc tyrosine is a useful building block for peptidesynthesis.

Using fluorosulfate-Fmoc tyrosine, several analogues of market peptidedrugs were synthesized with tyrosine-OSO₂F instead of tyrosine insequence by standard solid phase peptide synthesis methods that are wellknown in the art, demonstrating the utility of this functional group inpeptide chemistry.

Tyrosine O-sulfation is a common enzymatic post-translationalmodification that occurs while the secreted and transmembrane proteometraffics through the Golgi compartment of the cell. While it is clearthat phosphorylation and sulfation of tyrosine (Tyr) similarly modulateprotein-protein interactions and affect conformational changes within aprotein, much less is known about sulfation partly because sulfotyrosine(sY) peptides cannot be easily made by laboratories without substantialchemistry expertise.

Currently, several approaches are used for the solid-phase peptidesynthesis (SPPS) of sY-containing peptides, all of which have drawbacks.In one embodiment, the present invention provided an efficient approachto make sY-containing peptides wherein Fmoc-protected fluorosulfatedTyrosine (Y(OSO₂F)) is incorporated into the peptide-of-interest using anear-standard Fmoc solid phase synthesis strategy either manually or byusing a peptide synthesizer. Like other sulfur(VI) fluorides, aromaticfluorosulfates are hydrolytically stable, redox-resistant, and they donot serve as halogenation agents. They are very stable toward hydrolysisunder neutral or acidic conditions and are stable for up to two weeks inphosphate buffer at pH 10. However, they can become reactive in thepresence of an appropriate nucleophile under conditions that stabilizethe departure of the fluoride leaving group.

The ease of obtaining Fmoc protected Y(OSO2F) building block and thehigh stability of aromatic fluorosulfates enables the efficientsynthesis of peptides containing the Ar—O—SO₂F substructure using anFmoc chemistry strategy. The Fmoc-Y(OSO₂F)—OH (Fmoc-Y—SF, 1) used inSPPS is prepared in one step in high yield from the reaction ofcommercially available Fmoc-protected Tyr with sulforyl fluoride (gas)as described elsewhere herein (Scheme 1). Conveniently,2-methyl-piperidine (2-MP) was used to remove the Fmoc primary amineprotecting group during each SPPS coupling cycle (Scheme 2) instead ofpiperidine, as piperidine inefficiently reacts with the fluorosulfatefunctionality lowering the yield and purity of the desiredY(OSO₂F)-containing peptides. The fluorosulfate functional group isstable under the standard Rink amide resin-peptide cleavage(95:2.5:2.5=TFA:TIPS:H₂O) conditions used to liberate the side chaindeprotected peptide from the resin. Y(OSO₂F) residue(s) in thepeptide-of-interest were then converted into the sY functionality byhydrolysis with a base (Cs₂CO₃, or 1,8-diazabicyclo[5.4.0]undec-7-ene(DBU)) dissolved in ethylene glycol at 25° C. for 60-120 min withstirring (Scheme 3).

Five sY peptides 2-6 were prepared by an optimized protocol ofY(OSO₂F)-containing peptide synthesis and arylfluorosulfate hydrolysisemploying Cs₂CO₃/ethylene glycol (Table 4). The sequence of firstpeptide DADEsYL-NH₂ (P2; SEQ ID NO: 1) corresponds to a sequence in theepidermal growth factor receptor (EGFR) and P2 is expected to be a goodinhibitor of protein tyrosine phosphatase 1B. The sequence of amonosulfated peptide sYEsYLDsYDF-NH₂ (P3; SEQ ID NO: 2) and atrisulfated peptide sYEsYLDsYDF-NH₂ (P4; SEQ ID NO: 3) corresponds toresidue 5-12 of mature P-selectin glycoprotein ligand 1 (PGSL-1) thatbinds to P-selectin and plays important role in the rolling adhesion ofleukocytes on vascular endothelium. Disulfated peptideTTPDsYGHsYDDKDTLDLNTPVDK-NH₂ (P5; SEQ ID NO: 4) is from C5aR, aclassical G-protein coupled receptor that is implicated in manyinflammatory diseases. The sequence of tetrasulfated peptideDADSENSSFsYsYsYDsYLDEVAF-NH₂ (P6; SEQ ID NO: 5) corresponds to residue14-33 of chemokine receptor D6, which scavenges extracellularpro-inflammatory CC chemokines and suppresses inflammation andtumergenesis. Peptides P4-P6 contain multiple sY residues and theirsynthesis poses significant challenge to the coupling efficiency,stability and efficiency of hydrolysis for Y(OSO₂F) in differentsequence environments.

For all couplings in SPPS, including the coupling of amino acid 1, weused 5 equiv. of the appropriate side chain protected amino acidpreactivated with HCTU/HOBt/DIPEA for 30 min) (refs). The activatedamino acid was added to the resin-bound primary amine with shaking for acoupling period of 30-60 min to generate a new amide bond. Every Fmocprotecting group was removed employing 3 applications of 20% 2-MP indimethylformamide or N-methyl-2-pyrrolidone for 10 min. We used andprefer a 95:2.5:2.5=TFA:TIPS:H₂O deprotection solution (25° C., 180 min)to cleave the peptide-of-interest off the Rink resin and to liberate theside chain protecting groups, however the other cleavage/deprotectioncocktail mentioned above performs similarly. The crude yield of Y(OSO₂F)peptides carried out on a 200 μmol scale ranged from 77-89%. TheY(OSO₂F)-containing peptides can be easily purified by reversephase-high performance liquid chromatography (RP-HPLC), exemplified byDADEY(OSO₂F)L-NH₂ (P7; SEQ ID NO: 6) in a 64% yield. Fluorosulfatehydrolysis in peptide 7 was accomplished in ethylene glycol utilizingCs₂CO₃ as a base (10 equiv) and was followed by semi-preparative RP-HPLCusing a C18 column and an 20 mM ammonium acetate/CH₃CN mobile phasegradient, revealing near-quantitative conversion (FIG. 1A, S1 and S2).

In the optimization of the hydrolysis of Y(OSO₂F)-containing peptide 7to sY-containing peptide 2, we observed significant desulfation of sY inthe presence of base in aqueous solutions. Upon treating peptide 7 withCs₂CO₃ dissolved in methanol, we observed the apparent methylation ofcarboxylate side chains, presumably owing to the formation of aTyr-O—SO₂—OCH₃ intermediate. Although no consensus sequence is reachedfor tyrosine sulfation, acidic residues preponderate the known sites oftyrosine sulfation and such side reaction is likely to happen to othersulfated tyrosine sequences. (Lin 1992, Kehoe 2000, Seibert 2007,Teramoto 2013) While utilization of methanol/NH₃ (2M)/Cs₂CO₃ attenuatedmethylation, it was still observed. Utilizing Cs₂CO₃ dissolved inethanol resulted in peptide ethylation, consistent with formation ofTyr-O—SO₂—OCH₂CH₃ intermediate. With Cs₂CO₃ dissolved in isopropanol ortertiary butyl alcohol, no reaction occurred. Notably, whileCs₂CO₃/ethylene glycol and Cs₂CO₃/1,4 butanediol afforded quantitativehydrolysis with no side reactions, Cs₂CO₃/1,3 propanediol afforded <50%yield of sY peptide 2 and numerous side products.

Outside of hydrolysis methodology development explored in the precedingparagraph, there is no need to purify the crude Y(OSO₂F) peptidesresulting from 95:2.5:2.5=TFA:TIPS:H₂O treatment. Thus, the crudeY(OSO₂F)-containing peptides were directly subjected toarylfluorosulfate hydrolysis using ethylene glycol/Cs₂CO₃ method. Usingthis approach sY peptides P2-P6 were obtained in 36-67% yield (Table 4)after RP-HPLC purification using the column and conditions mentionedabove.

The synthesis of GDsYDSMKEPCFR-NH₂ (P8; SEQ ID NO: 7), a sY peptidecontaining Cys residue, is achieved by a different protocol employingDBU as the base in ethylene glycol. The sequence of 8 corresponds toresidue 19-30 of CXCR4, which is crucial for embryonic development andhave been implicated in cancer metastasis and HIV infection. sY has beenreported to make the largest contribution to CXCR4-CXCL12 binding. Usingthe optimized SPPS strategy and side chain deprotection/resin cleavagestrategy outlined above, the crude peptide GDY(OSO₂F)DSMKEPCFR-NH₂ (P9;SEQ ID NO: 8) (Table 4) was successfully synthesized. Peptide 9 was HPLCpurified in an isolated yield of 30% in order to optimize the hydrolysisstrategy for producing Cys-containing sY peptides. The standard ethyleneglycol/Cs₂CO₃ method didn't perform well because it appeared that wegenerated a Cys-S—COOH containing peptide. hydrolysis of peptide P9employing 5% DBU in ethylene glycol containing 0.5% dithiothreitol (DTT)was effective, however. This approach afforded sY peptide P8 in 25%isolated yield (Table 4). Adding DTT was the key to minimize theformation of byproducts.

The Fmoc synthesis of Y(OSO₂F) containing peptides described herein isboth practical and efficient. Standard side chain deprotection and resincleavage solutions perform well. Two different fluorosulfate hydrolysisprotocols are introduced for the efficient production of sY peptides.The method used depends on whether the peptide lacks or contains a Cysresidue. The facile synthesis described herein takes advantage of theunique reactivity of sulfur(VI) fluorides. This approach can easily beimplemented by commercial and academic peptide synthesis facilities.

TABLE 4 Isolated  Peptide Protein Sequence Yield SEQ ID NO: P2 EGFRDADEsYL-NH2 67% SEQ ID NO: 1 (988-993) P3 PGSL-1 YEsYLDYDF-NH2 54%SEQ ID NO: 2 (5-12) P4 PGSL-1 sYEsYLDsYDF-NH2 58% SEQ ID NO: 3 (5-12) P4C5aR TTPDsYGHsYDDKDTLDLNTPVDK-NH2 54% SEQ ID NO: 4 (7-28) P6 D6DADSENSSFsYsYsYDsYLDEVAF-NH2 36% SEQ ID NO: 5 (14-33) P7 EGFRDADEY(OSO2F)L-NH2 64% SEQ ID NO: 6 (988-993) P8 CXCR4 GDsYDSMKEPCFR-NH225% SEQ ID NO: 7 (19-30) P9 CXCR4 GDY(0502F)DSMKEPCFR-NH2 30%SEQ ID NO: 8 (19-30)

Four other commercial peptide drug derivatives were synthesized usingthe fluorosulfate-Fmoc tyrosine: Thymopentin LY001:RKDVYG*GG (SEQ ID NO:9); Oxytocin LY002:cCYIQNCPLG*GG (SEQ ID NO: 10); Arginine vasopressin(2 modified forms) LY003:CYFQNCPRG*GG (SEQ ID NO: 11); and IndolicidinLY005:I*GPWKWPYWPWRR-NH₂ (SEQ ID NO: 12) where Y is modified tyrosine(fluorosulfonated) and *G is propargylglycine.

Modified Oxytocin (SEQ ID NO: 10); Modified Indolicidin (SEQ ID NO: 12):

Modified Thymopentin (SEQ ID NO: 9); Modified Arginine Vasopressin (SEQID NO: 11):

The fluorosulfate ester groups of such peptides also can be converted tosulfate esters by selective hydrolysis with cesium carbonate and ammoniain methanol. For example, the modified indolicin was hydrolyzed to thecorresponding sulfate ester as verified by LCMS after direct transfer toPBS buffer.

Example 4. Fluorosulfate TTR Substrates

An ArOSO₂F functional group can serve as a covalent modifier in drugdesign. Almost 30% of the marketed drugs whose molecular targets areenzymes act by irreversible inhibition. J. Singh, R. C. Petter, T. A.Baille, A. Whitty, Nat. Rev. Drug Discov. 2011, 10, 307-317. Byinstalling an ArOSO₂F functional group on azo analogs of tafamidis, areversible stabilizer substrate of transthyretin (TTR), the analogs canbe transformed into an irreversible stabilizer. Small molecule analoguesof tafamidis react with TTR protein in PBS buffer (half-life is about 80min). As shown, below, installation of an-OSO₂F group via the generalprocedure for reactions of phenolic compounds with gaseous SO₂F₂(Ex.1(J)), above, changes the reversible parent inhibitor to irreversibleversions (i.e. azo compounds 4, 5, 6, and 7) which had half-livesranging from about 80 min to well over 6 hours after reaction with TTR.

Additionally, compounds in which the diazo group has been replaced by anoxadiazole have been prepared, as well. These compounds also exhibitedTTR binding activity with covalent binding to the active site, albeitwith subsequent hydrolysis by a lysine residue to form a sulfated lysinein the active site.

Example 5. Fluorosulfate Compounds for ¹⁸F PET Scan Applications

Due to its stability and ability to maintain affinity of the parentmolecule, ArOSO₂F is a useful compound for performing ¹⁸F PET scans forsmall molecules, peptides, and proteins. If a covalent reaction occurs,fast releasing fluoride ion will be readily confirmed by the enrichmentof ¹⁸F ion in bones, which would then be detectable by PET scantechniques. The reaction conditions are simple, fast, and provide fordirect loading of ¹⁸F on the target molecule. For example, a compoundwith a phenol group can be reacted with sulfuryl fluoride gas for about1-2 hours in buffer under mild condition to afford an Ar—OSO₂F compound.The very high conversion rate allows for simply removing the bufferwithout purification. Subsequent exchange of ¹⁹F by ¹⁸F can be readilyaccomplished by, e.g., exposure to Ag¹⁸F, or preferably ¹⁸F bifluoride.Proteins are an attractive target for this application as there iscurrently no known simple chemical way to install a small enough piececontaining a stable F molecule onto proteins. This process is similar toknown bioconjugation; however, this generally requires multiple stepsand larger molecules. For example, derivatization of dapagliflozin byreplacing the OEt group with an OSO₂F group provides a means ofaccessing an ¹⁸F version of dapagliflozin by ¹⁸F/¹⁹F exchange.

O-(2-[¹⁸F]fluoroethyl)-L-tyrosine has been used as a PET reagent in aclinical study. O-fluorosulfonyl-L-tyrosine is a fluorosulfonyl esteranalog of O-(2-fluoroethyl)-L-tyrosine, and is stable in HEK celllysates for at least 3 hours, which is a sufficient biostability for PETapplications. The ¹⁸F version of O-fluorosulfonyl-L-tyrosine can be usedfor PET imaging, and can be prepared rapidly and efficiently by exchangeof ¹⁹F by ¹⁸F by contacting the of non-radiolabeledO-fluorosulfonyl-L-tyrosine with ¹⁸F-enriched potassium bifluoride and apotassium complexing agent in a solvent such as acetonitrile.

Example 6. Fluorosulfate Compounds as “Click-Tag” Reagents and Probesfor Drug Discovery

Including an SO₂F group on a biologically active molecule can alsoprovide an opportunity to probe the active site of receptor molecules(e.g., by covalent reaction of the fluorosulfonyl moiety with an aminoacid side chain of a receptor when the fluorosulfonylated molecule isdocked in the active site). In this regard, the SO₂F group also can beconveniently combined with other functional probe/linking groups, forparticipation in additional coupling reactions with other usefulmaterials, such as dyes or other markers. For example, a molecule withboth an SO₂F group and an alkynyl group can be utilized as a couplingpartner in an azide/alkyne “click” coupling reaction. This processaffords a convenient and selective means of designing newfluorosulfonyl-based probes that include markers or other useful groupsattached to the biologically active drug structure.

In one example, ethynylestradiol fluorosulfate was readily reacted withan azo-substituted fluorescein derivative via a copper catalysedazide/alkyne complexing reaction (“click reaction”) to tether theestradiol derivative to fluorescein without affecting the OSO₂F group.

In the case of ethynylestradiol, the alkyne moiety already is part ofthe normal drug structure. Alternatively, an alkyne can be separatelyintroduced onto the drug structure in addition to the fluorosulfonylgroup, such as in the fluorosulfonylated propargyl vancomycin compounds,vancomycin-FS-1 and vancomycin-FS-2, from Example 2. Such probes canselectively pull down substrates for the biologically active portion ofthe probe from cell lysates and other complex mixtures of proteins andother biomolecules.

Ethynylestradiol fluorosulfate probes effectively capture tryptophanasefrom E coli lysates. The affinity of the ethynylestradiol fluorosulfateprobe described above for tryptophanase was as verified with recombinanttryptophanase, which was tagged with the probe with greater than 90%modification (50 micromolar tryptophanse, 500 micromolar probe, in TBSpH 8 buffer for 16 hours at 37° C.). Tryptophanase has been implicatedin biofilm formation, thus, the ethynylestradiol fluorosulfate materialsprovides a means for inhibiting biofilm formation.

Example 7. Covalent Attachment of SO₂F Compounds to Receptor Sites

The SO₂F moiety, in the various forms described herein also, in somecases, can provide a handle for covalent attachment of organic compoundsto receptor sites that include a nucleophilic amino acid side chain,such as a phenolic OH group, an amino group, a thiol, and the like, inthe reactive site in an orientation that can react to displace fluoridefrom sulfur. This concept is summarized in FIG. 21, which graphicallyillustrates docking of fluorosulfate and fluorosulfonyl-substitutedsubstrate molecules in a receptor site, and subsequent reaction todisplace fluoride and covalently bind the substrate molecule in theactive site. When several fluorosulfates or sulfonyl fluorides arepresent for potential reaction with the receptor, only molecules thathave an appropriate molecular configuration will interact/dock with theactive site of the receptor bind to the receptor site. Thus, thisprocess can be utilized as a screening assay, where a series of SO₂Fsubstituted candidate molecules (i.e., a library) are screened in afunctional assay for binding to a target receptor protein (see FIG. 22).

Example 8. Selective Sulfonation of Nucleophilic Amino Acid Side Chainsin Receptor Active Sites

In addition, the biologically active —OSO₂F compounds described hereincan be utilized in some instances to sulfonate a nucleophilic amino acidside chain in the active site of a receptor protein, e.g., by initialreaction of a receptor-docked fluorosulfate to displace fluoride, andsubsequent reaction with another nucleophilic side chain (Nu) todisplace the substrate molecule and form a Nu-SO₃— group, as illustratedin FIG. 23. In some cases, if the nucleophilic side chains both comprisethiols, an additional elimination step can occur to form a disulfidebond in the active site, as illustrated in FIG. 24.

Example 9. Sulfur(VI) Fluoride-Based Functional Groups on Probes thatSelectively and Covalently Modify Enzymes and Non-Enzymes in LivingCells

The binding-induced activation of sulfur (VI) fluoride functional groupsalso can be used in modifying proteins in biological systems. Aryl-SO₂Fand aryl-OSO₂F probes were studied in live HeLa cells using gel-basedassays and SILAC-based mass spectrometry approaches. Selective labelingof proteins by both aryl-SO₂F and aryl-OSO₂F probes were observed andthe protein targets were identified.

We examined the reactivity and selectivity of covalent labeling byaryl-SO₂F and aryl-OSO₂F probes and select probes SF-3 and OSF-2 fortarget identification via the SILAC technique (FIG. 25). Target proteinsfor both probes were found to befunctional unrelated enzymes ornon-enzymes. Surprisingly, two non-enzymes, FABP5 and CRABP2, werelabeled in both cases. Using recombinant FABP5 and CRABP2 the labelingevent was confirmed and the site of labeling was identified as afunctionally important tyrosine in an Arg-Tyr-Arg cluster that couldbind the carboxyl group in fatty acids (FIG. 26). Since the expressionsof these intracellular lipid-binding proteins (iLBPs) are quite tissuespecific it was hypothesized that other iLBPs with this structuralfeature could be labeled by SF-3 and OSF-2. Such labeling was confirmedusing recombinant FABP3 and FABP4.

Covalent inhibitors based on SF-3 and OSF-2 (similar molecules withoutan alkyne handle) were synthesized and used to compete with the probes(FIGS. 27(A and B)). With increasing concentrations of covalentinhibitors a decreasing labeling event toward FABP5/CRABP2 was observed,suggesting the labeling is chemoselective and could be completed atcertain concentration. These SO₂F and OSO₂F probes also were used toexamine known and previously unknown non-covalent inhibitors of iLBPs inlive HeLa cells (FIG. 27(C)). Surprisingly, an SOAT inhibitor (AvasimibePfizer, phase III) could compete out the labeling on FABP5 and CRABP2 inlive HeLa cells suggesting strong binding in the substrate bindingpockets. These studies provide a new way to covalently target iLBPs andsuggest that the promiscuous reactivity of sulfur (VI) fluoridefunctional groups can be tuned and utilized for selective proteinmodification.

Example 10. Additional Examples of ESF and SO₂F₂ Modified Drugs andOther Biologically Active Compounds

Biologically active compounds, such as drugs, enzyme inhibitors, othertherapeutic agents, agrochemicals (e.g., herbicides, fungicides, andpesticides), and the like, which have a pendant primary or secondaryamino nitrogen group are readily reactive toward ethylenesulfonylfluoride (ESF), as described in detail herein, to form an ESF derivativevia Michael addition of the amino group to the ESF double bond. In thecase of primary amino compounds, one or two ESF groups can be added, bycontrolling the stoichiometry (one equivalent of ESF will replace onehydrogen of a primary amine with an fluorosulfonylethyl group; if twoequivalents of ESF are used, both hydrogens of the amino group will bereplaced by fluorosulfonylethyl groups). The reaction with ESF can becarried out in the presence of hydroxyl groups, including phenolichydroxyl groups, without any substantial interference. FIGS. 28 to 35provide examples of biologically active compounds that can be reactedwith ESF to form ESF adducts according to the methods described herein.

Biologically active compounds that have one or more pendant aromatic orheteroaromatic hydroxyl group or a pendant secondary amino group arereadily reactive toward SO₂F₂ in the presence of a base (e.g., atertiary amine) to form fluorosulfate esters with the hydroxyl group, ora fluorosulfamate with the amino group, as described in detail herein.FIGS. 36 and 37 provide examples of biologically active compounds thatcan be reacted with SO₂F₂ in the presence of a base to formfluorosulfates and fluorosulfamides, according to the methods describedherein.

Methods for evaluating the activity of the various modified biologicallyactive compounds described herein are well known in the art, as most ofthe biologically active core compounds from which thefluorosulfonyl-containing derivatives (e.g., fluorosulfates,fluorosufamates, and ESF adducts, collectively referred to asSF-modified compounds) are prepared have been extensively studied in theliteratue and many are or have been commercial drugs or products.Non-limiting examples of methods for evaluating the activity of some ofthe SF-modified compounds described herein are based on methods forassaying the activity of the parent compounds, as described in theparagraphs below.

Mephentermine is obtained from Cerilliant (Saint Louis, Mo.). TheSF-modified Mephentermine is evaluated for activity in assay asdescribed by G. Fawaz and J. Simaan, “The Tachyphylaxis caused bymephentermine and tyramine,” British Journal of Pharmacology, Vol 24(1965) pp. 526-531.

Mecamylamine hydrochloride is obtained from Sigma-Aldrich® (Milwaukee,Wis.). The SF-modified mecamylamine is evaluated for activity in assayas described by N. Gentile, et al., “Sexually diergichypothalamic-pituitary-adrenal (HPA) responses to single-dose nicotine,continuous nicotine infusion, and nicotine withdrawal by mecamylamine inrats,” Brain Research Bulletin, Vol 85 (2011) pp. 145-152.

Levallorphan tartrate salt is obtained from Sigma-Aldrich® (Milwaukee,Wis.). The SF-modified levallorphan compound is evaluated for activityin assay as described by B. Brdar and P. Fromageot, “Inhibition of viralRNA synthesis by levallorphan,” FEBS Letters, Vol. 6, No. 3 (1970) pp.190-192.

Naltrexone Hydrochloride is obtained from Sigma-Aldrich® (Milwaukee,Wis.). The SF-modified naltrexone compound is evaluated for activity inassay as described by C. Moore, “The efficacy of a low dose combinationof topiramate and naltrexone on ethanol reinforcement and consumption inrat models,” Pharmacology, Biochemistry and Behavior, Vol. 116 (2014)pp. 107-115.

Levothyroxine is obtained from Sigma-Aldrich® (Saint Louis, Mo.). TheSF-modified compound is evaluated for activity in assay as described byD. Pabla, et al., “Intestinal permeability enhancement of levothyroxinesodium by straight chain fatty acids studied in MDCK epithelial cellline,” European Journal of Pharmaceutical Sciences, Vol. 40 (2010) pp.466-472.

Liothyronine is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified liothyronine compound is evaluated for activity in assay asdescribed by S. Wu, et al., “Tissue responses to thyroid hormone in akindred with resistance to thyroid hormone harboring a commonlyoccurring mutation in the thyroid hormone receptor β gene (P453T),”Journal of Laboratory and Clinical Medicine, Vol. 146, Issue 2 (2005)pp. 85-94.

Metaraminol is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified metaraminol compound is evaluated for activity in assay asdescribed by A. Sagie, et al., “Effect of metaraminol during acuteinferior wall myocardial infarction accompanied by hypotension:preliminary study,” Journal of the American College of Cardiology, Vol.10, Issue 5 (1987) pp. 1139-1144.

Nabilone is obtained from Sigma-Aldrich® (Saint Louis, Mo.). TheSF-modified nabilone compound is evaluated for activity in assay asdescribed by J. Lile, et al., “Separate and combine effects of thecannabinoid agonists nabilone and A9-THC in humans discriminatingA9-THC,” Drug and Alcohol Dependence, Vol. 116, Issues 1-3 (2011) pp.86-92.

Sulfadoxine is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified compound is evaluated for activity in assay as described byC. Happi, et al., “Polymorphisms in Plasmodium falciparum: dhfr and dhpsgenes and age related in vivo sulfadoxine-pyrimethamine resistance inmalaria-infected patients from Nigeria” Acta Tropica, Vol. 95 (2005) pp.183-193.

Sumatriptan is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified sumatriptan compound is evaluated for activity in assay asdescribed by Y. Watanabe, et al., “Monitoring cortical hemodynamicchanges after sumatriptan injection during migraine attack bynear-infrared spectroscopy,” Neuroscience Research, Vol. 69 (2011) pp.60-66.

Tacrine is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified tacrine compound is evaluated for activity in assay asdescribed by C. Gao, et al., “Tacrine induces apoptosis throughlysosome- and mitochondria-dependent pathway in HepG2 cells,” ToxicologyIn Vitro, Vol. 28, Issue 4 (2004) pp. 667-674.

Theophyline is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified theophyline compound is evaluated for activity in assay asdescribed by E. Hashimoto, et al., “Adenosine as an endogenous mediatorof hypoxia for induction of vascular endothelial growth factor mRNA inU-937 cells,” Biochemical and Biophysical Research Communications, Vol.204, No. 1 (1994) pp. 318-324.

Tadalafil is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified tadalafil compound is evaluated for activity in assay asdescribed by C. Zhu, et al., “Preventive effect of phosphodiesterase 5inhibitor Tadalafil on experimental post-pyelonephritic renal injury inrats,” Journal of Surgical Research, Vol. 186 92014) pp. 253-261.

Tranexamic is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified tranexamic compound is evaluated for activity in assay asdescribed by H. Kakiuchi, et al., “Tranexamic acid induces kaolin intakestimulating a pathway involving tachykinin neurokinin 1 receptors inrats,” European Journal of Pharmacology, Vol. 723 (2014) pp. 1-6.

Varenicline is obtained from Sigma-Aldrich® (St. Louis, Mo.).SF-Modified varenicline compound is evaluated for activity in assay asdescribed by C. Cunningham and L. McMahon, “The effects of nicotine,varenicline, and cystine on schedule-controlled responding in mice:Differences in α4β2 nicotinic receptor activation,” European Journal ofPharmacology, Vol. 654 (2011) pp. 47-52.

Vancomycin is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified compound is evaluated for activity in assay as described byT. Dilworth, et al., “Vancomycin and piperacillin-tazobactam againstmethicillin-resistant Staphylococcus aureus and vancomycin-intermediateStaphylococcus aureus in an in vitro pharmacokinetic/pharmacodynamicmodel,” Clinical Therapeutics, Vol. 36 (2014) pp. 1335-1344.

Vigabatrin is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified compound is evaluated for activity in assay as described byJ. Plum, et al., “The anti-epileptic drug substance vigabatrin inhibitstaurine transport in intestinal and renal cell culture models,”International Journal of Pharmaceutics, Vol. 473 (2014) pp. 395-397.

Salicyclic Acid is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified compound is evaluated for activity in assay as described byH. Chen, et al., “Salicylic acid mediates alternative signaltransduction pathways for pathogenesis-related acidic β-1,3-glucanase(protein N) induction in tobacco cell suspension culture,” Journal ofPlant Physiology, Vol. 159 (2002) pp. 331-337.

Terbutaline is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified compound is evaluated for activity in assay as described byA. Hodi, et al., “Tocopherol inhibits the relaxing effect of terbutalinein the respiratory and reproductive tracts of the rat: The role of theoxidative stress index,” Life Sciences, Vol 105 (2014) pp. 48-55.

Rotigotine is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified compound is evaluated for activity in assay as described byS. Oster, et al., “Rotigotine protects against glutamate toxicity inprimary dopaminergic cell culture,” European Journal of Pharmacology,Vol. 724 (2014) pp. 31-42.

Prazosin hydrochloride is obtained from Alfa Aesar® (Ward Hill, Mass.).The SF-modified Prazosin compound is evaluated for activity in assay asdescribed by A. Antonello, et al., “Design, synthesis, and biologicalevaluation of prazosin-related derivatives as multipotent compounds,”Journal of Medicinal Chemistry, Vol. 48, No. 1 (2005), pp. 28-31.

Pregabalin is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified Pregabalin compound is evaluated for activity in assay asdescribed by K. Fink, et al., “Inhibition of neuronal Ca2+ influx bygabapentin and pregabalin in the human neocortex,” Neuropharmacology,Vol. 42 (2002) pp. 229-236.

Procainamide is obtained from Alfa Aesar® (Ward Hill, Mass.). TheSF-modified procainamide compound is evaluated for activity in assay asdescribed by B. Lee, et al., “Procainamide is a specific inhibitor ofDNA methyltransferase 1,” The Journal of Biological Chemistry, Vol. 280,No. 49 (2005) pp. 40749-40756.

Procarbazine is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified procarbazine compound is evaluated for activity in assay asdescribed by D. Clive, et al., “Procarbazine is a potent mutagen at theheterozygous thymidine kinase (tk+/−) locus of mouse lymphoma assay,”Mutagenesis, Vol. 3, No. 2 (1988) pp. 83-87.

Propafenone is obtained from Alfa Aesar® (Ward Hill, Mass.). TheSF-modified propafenone compound is evaluated for activity in assay asdescribed by H. Komura and M. Iwaki, “Nonlinear pharmacokinetics ofpropafenone in rats and humans: application of a substrate depletionassay using hepatocytes for assessment of nonlinearity,” Drug Metabolismand disposition, Vol. 33 (2005), pp. 726-732.

Protriptyline is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified protriptyline compound is evaluated for activity in assay asdescribed by S. Bansode, et al., “Molecular investigations ofprotriptyline as a multi-target directed ligand in alzheimer's disease,”PLoS ONE, Vol. 9, Issue. 8 (2014) e105196.doi:10.1371/journal.pone.0105196.

Pseudoephedrine is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified pseudoephedrine compound is evaluated for activity in assayas described by Z. Wu, et al., “Pseudoephedrine/ephedrine shows potentanti-inflammatory activity against TNF-α-mediated acute liver failureinduced by lipopolysaccharide/D-galactosamine,” European Journal ofPharmacology, Vol. 724 (2014), pp. 112-121.

Ramipril is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified ramipril compound is evaluated for activity in assay asdescribed by X. Ji, et al., “Comparison of cardioprotective effectsusing ramipril and DanShen for the treatment of acute myocardialinfarction in rats,” Life Sciences, Vol. 72 (2003) pp. 1413-1426.

Rasagiline is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified rasagiline compound is evaluated for activity in assay asdescribed by Y. Aluf, et al., “Selective inhibition of monoamine oxidaseA or B reduces striatal oxidative stress in rats with partial depletionof the nigro-striatal dopaminergic pathway,” Neropharmacology, Vol. 65(2013) pp. 48-57.

Reboxetine is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified reboxetine compound is evaluated for activity in assay asdescribed by B. Benedetto, et al., “N-desalkylquetiapine activatesERK1/2 to induce GDNF release in C6 glioma cells: A putative cellularmechanism for quetiapine as antidepressant,” Neuropharmacology, Vol. 62(2012) pp. 209-216.

Rimantadine is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified rimantadine compound is evaluated for activity in assay asdescribed by G. Stamatiou, et al., “Heterocyclic rimantadine analogueswith antiviral activity,” Bioorganic & Medicinal Chemistry, Vol. 11(2003) pp. 5485-5492.

Ritodrine is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified ritodrine compound is evaluated for activity in assay asdescribed by F. Plenge-Tellechea, et al., “Ritodrine inhibition of theplasma membrane Ca2+-ATPase from human erythrocyte,” Archives ofBiochemistry and Biophysics, September 15, Vol. 357, No. 2 (1998) pp.179-184.

S-adenosylmethionine is obtained from Sigma-Aldrich® (St. Louis, Mo.).The SF-modified s-adenosylmethionine compound is evaluated for activityin assay as described by F. Zhang, et al., “S-adenosylmethionineinhibits the activated phenotype of human hepatic stellate cells viaRac1 and Matrix metalloproteinases,” International Immunopharmacology,Vol. 19 (2014) pp. 193-200.

Salmeterol is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified salmeterol compound is evaluated for activity in assay asdescribed by Andrea Teschemacher and Horst Lemoine, “Kinetic analysis ofdrug-receptor interactions of long-acting 32 sympathomimetics inisolated receptor membranes: evidence against prolonged effects ofsalmeteroland formoterol on receptor-coupled adenylyl cyclase,” TheJournal of Pharmocology and Experimental Therapeutics, Vol. 288, No. 3(1999) pp. 1084-1092.

Saxagliptin is obtained from Astatech Inc. (Bristol, Pa.). TheSF-modified saxagliptin compound is evaluated for activity in assay asdescribed by J. Kosaraju, et al., “Saxagliptin: a dipeptidyl peptidase-4inhibitor ameliorates streptozotocin induced Alzheimer's disease,”Neuropharmacology, Vol 72 (2013) pp. 291-300.

Sitagliptin is obtained from Astatech Inc. (Bristol, Pa.). TheSF-modified sitagliptin compound is evaluated for activity in assay asdescribed by Tremblay, A., “Effects of sitagliptin therapy on markers oflow-grade inflammation and cell adhesion molecules in patients with type2 diabetes,” Metabolism Clinical and Experimental, Vol 63 (2014) pp.1131-1148.

Sparfloxacin is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified sparfloxacin compound is evaluated for activity in assay asdescribed by E. Efthimiadou, et al., “Mononuclear dioxomolybdenum (VI)complexes with the quinolones enrofloxacin and sparfloxacin: Synthesis,structure, antibacterial activity and interaction with DNA,” Polyhedron,Vol. 27 (2008) pp. 349-356.

Gabapentin is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified gabapentin compound is evaluated for activity in assay asdescribed by F. Kilic, et. al., “Antinociceptive effects of gabapentin &its mechanism of action in experimental animal studies,” Indian J. Med.Res., May; 135(5) (2012) pp. 630-635.

Sertraline hydrochloride is obtained from Sigma-Aldrich® (Milwaukee,Wis.). The SF-modified sertraline compound is evaluated for activity inassay as described by R. Vijaya and K. Ruckmani, “In vitro and In vivocharacterization of the transdermal delivery of sertraline hydrochlorideFilms,” Journal of Pharmaceutical Sciences, Vol. 19, No. 6 (2011) pp.424-432.

Lisinopril is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified lisinopril compound is evaluated for activity in assay asdescribed by C. Constantinescu, et. al., “Catopril and lisinoprilsuppress production of interleukin-12 by human peripheral bloodmononuclear cells,” Immunology Letters, 62 (1998) pp. 25-31.

Amphetamine is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified amphetamine compound is evaluated for activity in assay asdescribed by T. Kanbayashi, et. al., “Implication of dopaminergicmechanisms in the wake-promoting effects of amphetamine: A study of D-and L-derivatives in canine narcolepsy,” Neuroscience, Vol. 99, No. 4(2000) pp. 651-659.

Fluoxetine is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified fluoxetine compound is evaluated for activity in assay asdescribed by M. Bianchi, et. al., “Effects of chlomipramine andfluoxetine on subcutaneous carrageenin-induced inflammation in the rat,”Inflammation Research, Vol. 44 (1995), pp. 466-469.

Bupropion is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified bupropion compound is evaluated for activity in assay asdescribed by S. Learned-Coughlin, “In vivo activity of bupropion at thehuman dopamine transporter as measured by positron emission tomography,”Biological Psychiatry, Vol. 54 (2003), pp. 800-805.

Nadolol is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified nadolol compound is evaluated for activity in assay asdescribed by W. Wu and S. Pruett, “Suppression of splenic natural killercell activity in a mouse model for binge drinking, II. Role of theneuroendocrine system,” The Journal of Pharmacology and ExperimentalTherapeutics, 278 (1996) pp. 1331-1339.

Albuterol sulfate is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified albuterol compound is evaluated for activity in assay asdescribed by J. Cancado, et. al., “Effect of airway acidosis andalkalosis on airway vascular smooth muscle responsiveness to albuterol,”BMC Pharmacology and Toxicology, (2015) 16:9.

Phentermine is obtained from Sigma-Aldrich® (Round Rock, Tex.). TheSF-modified phentermine compound is evaluated for activity in assay asdescribed by J. Kang, et. al., “Randomized controlled trial toinvestigate the effects of a newly developed formulation of phenterminediffuse-controlled release for obesity,” Diabetes, Obesity andMetababolism, Vol. 12 (2010) pp. 876-882.

Atenolol is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified atenolol compound is evaluated for activity in assay asdescribed by S. Dey, et. al., “Formulation and evaluation of fixed-dosecombination of bilayer gastroretentive matrix table containingatorvastatin as fast-release and atenolol as sustained-release,” BiomedResearch International, Volume 2014, Article ID 396106, 12 pages.

Cefadroxil is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified cefadroxil compound is evaluated for activity in assay asdescribed by X. Chen, et. al., “Effect of transporter inhibition on thedistribution of cefadroxil in rat brain,” Fluid Barriers of the CNS,(2014) 11:25.

Warfarin is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified warfarin compound is evaluated for activity in assay asdescribed by T. Li, et. al., “Identification of the gene for vitamin Kepoxide reductase,” Nature, Vol. 427 (2004) p. 541-543.

Butorphanol is obtained from Sigma-Aldrich® (St. Louis, Mo.). TheSF-modified hydromorphone compound is evaluated for activity in assay asdescribed by S. Walsh, et. al., “Enadoline, a selective kappa opioidagaonist: comparison with butorphanol and hydromorphone in humans,”Psychopharmacology, Vol. 157 (2001) pp. 151-162.

Hydromorphone hydrochloride is obtained from Sigma-Aldrich® (Milwaukee,Wis.). The SF-modified hydromorphone compound is evaluated for activityin assay as described by S. Walsh, et. al., “Enadoline, a selectivekappa opioid agaonist: comparison with butorphanol and hydromorphone inhumans,” Psychopharmacology, Vol. 157 (2001) pp. 151-162.

Estradiol is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified estradiol compound is evaluated for activity in assay asdescribed by V. Pentikainen, et. al., “Estradiol acts as a germ cellsurvival factor in the human testis in vitro,” The Journal of ClinicalEndocrinology & Metabolism, Vol. 85, Vol. 5 (2000) pp. 2057-2067.

Indolicidin is obtained from AnaSpec, Inc. (Fremont, Calif.). TheSF-modified indolicidin compound is evaluated for activity in assay asdescribed by Selsted, et. al., “Indolicidin, a Novel BactericidalTridecapeptide Amide from Neutrophils,” The Journal of BiologicalChemistry, Vol. 267, No. 7, Issue of March 5 (1992) pp. 4292-4295.

Thymopentin is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified thymopentin compound is evaluated for activity in assay asdescribed by Fan, et. al., “Thymopentin (TP5), an immunomodulatorypeptide, suppresses proliferation and induces differentiation in HL-60cells,” Biochimica et Biophysica Acta, Vol. 1763 (2006) pp. 1059-1066.

Oxytocin is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified oxytocin compound is evaluated for activity in assay asdescribed by U.S. Pharmacopeia Pharmacopeial Forum: Volume No. 29(6) p.1946.

Arginine Vasopressin is obtained from Sigma-Aldrich® (Milwaukee, Wis.).The SF-modified arginine vassopressin compound is evaluated for activityin assay as described by U.S. Pharmacopeia Pharmacopeial Forum: VolumeNo. 29(6)31(4) p. 1127.

Tetrahydrocannabinol is obtained from Sigma-Aldrich® (Saint Louis, Mo.).The SF-modified tetrahydrocannabinol compound is evaluated for activityin assay as described by M. Parolini and A. Binelli, “Oxidative andgenetic responses induced by A9-Tetrahydrocannabinol (Δ-9-THC) toDreissena polymorpha,” Science of the Total Environment, Vol. 468-469(2014) pp. 68-76.tette

Methylphenidate is obtained from Sigma-Aldrich® (Saint Louis, Mo.). TheSF-modified methylphenidate compound is evaluated for activity in assayas described by A. Issy and E. Del Bel, “7-Nitroinadazole blocks theprepulse inhibition disruption and c-Fos increase induced bymethylphenidate,” Behavioural Brain Research, Vol. 262 (2014) pp. 74-83.

Desloratadine is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified desloratadine compound is evaluated for activity in assay asdescribed by Y. Lin, et al., “Design, synthesis and biological activityevaluation of desloratadine analogues as H1 receptor antagonists,”Bioorganic 7 Medicinal Chemistry, Vol. 21 (2013) pp. 4178-4185.

Anisomycin is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified anisomycin compound is evaluated for activity in assay asdescribed by X. Guo, et al., “Epigenetic mechanisms of amyloid-βproduction in anisomycin-treated SH-SY5Y cells,” Neuroscience, Vol. 194(2011) pp. 272-281.

Strobilurin F is isolated as described by A. Fredenhagen, et al.,“Strobilurins F, G and H, three new antifungal metabolites from BolineauLutea I. fermentation, isolation and biological activity,” The Journalof Antibiotics, Vol. XLIII, No. 6 (1990) pp. 655-660. The SF-modifiedstrobilurin compound is evaluated for activity in assay as described byJ. Sudisha, et al., “Comparative efficacy of strobilurin fungicidesagainst downy mildew disease of pearl millet,” pesticide biochemistryand Physiology, Vol. 81 (2005) pp. 188-197).

Cyclopamine is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified cyclopamine compound is evaluated for activity in assay asdescribed by T. Takahasi, et al., “cyclopamine induces eosinophilicdifferentiation and upregulates CD44 expression in myeloid leukemiacells,” Leukemia Research, Vol. 35 (2011) pp. 638-645.

Capsaicin is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified capsaicin compound is evaluated for activity in assay asdescribed by R. Terayama, et al., “Assessment of intraoral mucosal paininduced by the application of capsaicin,” Oral Biology, Vol. 59 (2014)pp. 1334-1341.

Trifloxystrobin is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified trifloxystrobin compound is evaluated for activity in assayas described by B. Zhu, et al., “Assessment of trifloxystrobin uptakekinetics, developmental toxicity and mRNA expression in rare minnowembryos,” Chemosphere, Vol. 120 (2015) pp. 447-455.

Impidacloprid is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified impidacloprid compound is evaluated for activity in assay asdescribed by X. Yang, et al., “Two cytochrome P450 genes are involved inimidacloprid resistance in field populations of the whitefly, Bemisiatabaci, in China,” Pesticide Biochemistry and Physiology, Vol. 107,Issue 3 (2013) pp. 343-350.

Acetamiprid is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified acetamiprid compound is evaluated for activity in assay asdescribed by T. Cavas, et al., “In vitro genetoxicity evaluation ofacetamiprid in CaCo-2 cells using the micronucleus, comet and y¹H2AXfoci assays,” Pesticide Biochemistry and Physiology, Vol. 104 (2012) pp.212-217.

Nitenpyram is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified nitenpyram compound is evaluated for activity in assay asdescribed by T. Perry, et al., “Effects of mutations in Drosophilanicotinic acetylcholine receptor subunits on sensitivity to insecticidestargeting nicotinic acetylcholine receptors,” Pesticide Biochemistry andPhysiology, Vol. 102 (2012) pp. 56-60.

Fipronil is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified fipronil compound is evaluated for activity in assay asdescribed by C. Baker, et al., “Efficacy of a novel topical combinationof fipronil, (S)-methoprene, eprinomectin and praziquantel against adultand immature stages of the cat flea (Ctenocephalides felis) on cats,”Veterinary Parasitology, Vol. 202 (2014) p. 54-58.

Lufenuron is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified lufenuron compound is evaluated for activity in assay asdescribed by M. Breijo, et al., “An insect growthinhibitor—lufenuron—enhances albendazole activity against hydatid cyst,”Veterinary Parasitology, Vol. 181 (2011) pp. 341-344.

Fluconazole is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified fluconazole compound is evaluated for activity in assay asdescribed by Q. Yu, et al., “In vitro activity of verapamil alone and incombination with fluconazole or tunicamycin against Candida albicansbiofilms,” International Journal of Antimicrobial Agents, Vol. 41 (2013)179-182.

Nitroxoline (8-hydroxy-5-nitroquinoline) is obtained from Sigma-Aldrich®(Milwaukee, Wis.). The SF-modified nitroxoline compound is evaluated foractivity in assay as described by G. Murugasu-Oei and T. Dick, “In vitroactivity of the chelating agents nitroxoline and oxine againstMycobacterium bovis BCG,” International Journal of Antimicrobial Agents,Vol. 18 (2001) pp. 579-582.

Pentazocine is obtained from Sigma-Aldrich® (Saint Louis, Mo.). TheSF-modified pentazocine compound is evaluated for activity in assay asdescribed by P. Martin, et al., “The sigma receptor ligand(+)-pentazocine prevents apoptotic retinal ganglion cell death inducedin vitro by homocysteine and glutamate,” Molecular Brain Research, Vol.123, Issues 1-2 (2004) pp. 66-75.

Isocarboxazid is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified isocarboxazid compound is evaluated for activity in assay asdescribed by A. Klegeris and P. McGeer, “R-(−)-Deprenyl inhibitsmonocytic THP-1 cell neurotoxicity independently of monoamine oxidaseinhibition,” Experimental Neurology, Vol. 166 (2000) pp. 458-464.

Indapamide is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified indapamide compound is evaluated for activity in assay asdescribed by C. Ren, et al., “Design and in vivo evaluation of anindapamide transdermal patch,” International Journal of Pharmaceutics,Vol. 370 (2009) pp. 129-135.

Ketamine is obtained from Sigma-Aldrich® (Round Rock, Tex.). TheSF-modified ketamine compound is evaluated for activity in assay asdescribed by G. Vasconcelos, et al., “Alpha-lipoic acid alone andcombined with clozapine reverses schizophrenia-like symptoms induced byketamine in mice: participation of antioxidant, nitrergic andneurotrophic mechanisms,” Schizophrenia Research, Vol. 165 (2015) pp.163-170.

Lomefloxacin is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified lomefloxacin compound is evaluated for activity in assay asdescribed by Y. Zhou, et al., “Synthesis, cytotoxicity and topoisomeraseII inhibitory activity of lomefloxacin derivatives,” Bioorganic &Medicinal Chemistry Letters, Vol. 23 (2013) pp. 2974-2978.

Moxifloxacin is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified moxifloxacin compound is evaluated for activity in assay asdescribed by F. Hurtado, et al., “Enhanced penetration of moxifloxacininto rat prostate tissue evidenced by microdialysis,” InternationalJournal of Antimicrobial Agents, Vol. 44, Issue 4 (2014) pp. 327-333.

Paroxetine is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified paroxetine compound is evaluated for activity in assay asdescribed by Y.

Sugimoto, et al., “Involvement of the sigma receptor in theantidepressant-like effects of fluvoxamine in the forced swimming testin comparison with the effects elicited by paroxetine,” European journalof Pharmacology, Vol. 696, Issues 1-3 (2012) pp. 96-100.

Methazolamide is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified methazolamide compound is evaluated for activity in assay asdescribed by M. Corena, et al., “Degradation and effects of thepotential mosquito larvicides methazolamide and acetazolamide insheepshead minnow (Cyprinodon variegates),” Ecotoxicology andEnvironmental Safety, Vol. 64 (2006) pp. 369-376.

Methylphenidate is obtained from Sigma-Aldrich® (Saint Louis, Mo.). TheSF-modified methylphenidate compound is evaluated for activity in assayas described by C. Wrenn, et al., “Effects of clonidine andmethylphenidate on motor activity in Fmr1 knockout mice,” NeuroscienceLetters, Vol. 585 (2015) pp. 109-113.

Milnacipran is obtained from Sigma-Aldrich® (Round Rock, Tex.). TheSF-modified milnacipran compound is evaluated for activity in assay asdescribed by M. Yamauchi, et al., “A combination of mirtazapine andmilnacipran augments the extracellular levels of monoamines in the ratbrain,” Neuropharmacology, Vol. 62 (2012) pp. 2278-2287.

Maprotiline is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified maprotiline compound is evaluated for activity in assay asdescribed by C. Jan, et al., “Mechanism of maprotiline-inducedapoptosis: Role of [Ca2+]I, ERK, JNK and caspase-3 signaling pathways,”Toxicology, Vol. 304 (2013) 1-12.

Nortriptyline is obtained from Sigma-Aldrich® (Milwaukee, Wis.). TheSF-modified nortriptyline compound is evaluated for activity in assay asdescribed by C. Piubelli, et al., “Nortriptyline influences proteinpathways involved in carbohydrate metabolism and actin-related processesin a rat gene-environment model of depression,” EurpoeanNeuropsycholpharmacology, Vol. 21, Issue 7 (2011) pp. 545-562.

In addition, a fluorosulfate analog of the drug Riluzole, which is usedto treat amyotrophic lateral sclerosis, can be prepared by reaction ofcommercially available 2-amino-6-hydroxy-benzothiazole with SO₂F₂ in thepresence of a base (e.g., triethylamine), as shown below.

Example 11. Inhibitors of Soluble Epoxidase Hydrolase (sEH)

Soluble epoxide hydrolase (sEH) is a bifunctional, homodimeric enzymewith hydrolase and phosphatase activity, sEH is highly expressed in theliver, but it is also expressed in tissues such as vascular endothelium,leukocytes, red blood cells, smooth muscle cells, adipocytes, as well asthe kidney proximal tubule. sEH metabolizes cis-epoxyeicosatrienoicacids (EETs) as well as other lipid mediators, and as such sEH plays arole in several diseases including hypertension, cardiac hypertrophy,arteriosclerosis, brain and heart ischemia injury, cancer and pain.Fluorosulfonyl derivatives of soluble epoxide hydrolase (sEH) inhibitorsare useful for treatment of sEH-mediated diseases or conditions.

A. Synthesis of(S)-4-(3-(1-(2-methylbutanoyl)piperidin-4-yl)ureido)phenylsulfurofluoridate

4-Aminophenyl sulfurofluoridate (also known as aniline-4-fluorosulfate;80 mg, 419 μmol, 1.0 eq) and triethyl amine (Et₃N; 46.5 mg, 461 μmol,1.1 eq) were dissolved in CH₂Cl₂ (5 mL) with stirring at −78° C.Triphosgene (46 mg, 155 μmol, 0.37 eq) dissolved in CH₂Cl₂ (5 mL) wasadded dropwise at −78° C. The reactants were then warmed to 0° C. andstirred for 30 min. Thereafter the reactants and reaction products werecooled to 0° C. (S)-1-(4-aminopiperidin-1-yl)-2-methylbutan-1-one (84mg, 461 μmol, 1.2 eq) and Et₃N (46.5 mg, 461 μmol, 1.1 eq) dissolved inCH₂Cl₂ (DCM; 5 mL) were added slowly and the resulting reaction mixturewas further stirred at room temperature for 12 h. The reaction was thenquenched with the addition of HCl solution (1M, 15 mL). An organic layerwas collected from the reaction mixture and the remaining aqueous layerwas further extracted with EtOAc three times. The obtained organiclayers were combined and washed with saturated NaCl solution. The washedorganic layer was dried over anhydrous magnesium sulfate and wasconcentrated under vacuum. The obtained product (85 mg; 50.6%) waseluted by flash chromatography with (EtOAc:Hexane/7:3). The product wasfurther purified by crystallization (MeOH with water). Yield 50.6%. ¹HNMR (d₆-DMSO, 300 Mhz): ∂ 0.80-0.90 (m, 3H), 0.97 (t, J=5 Hz, 3H),1.2-1.4 (m, 3H), 1.4-1.6 (m, 1H), 1.7-1.9 (m, 2H), 2.7-2.9 (m, 2H), 3.16(t, J=12 Hz, 1H), 3.6-3.8 (m, 1H), 3.88 (d, J=12.6 Hz, 1H), 4.21 (br,1H), 6.32 (t, J=7.5 Hz, 1H), 7.43 (d, J=9 Hz, 2H), 7.54 (d, J=9 Hz, 2H),8.68 (d, J=8 Hz, 1H); Melting point (° C.): 186.5-188.0 (187.3).

B. Synthesis of 4-(3-(1-propionylpiperidin-4-yl)ureido)phenylSulfurofluoridate

B (a). 26 mg (0.1 mmol scale) tert-butyl(1-propionylpiperidin-4-yl)carbamate dissolved in 0.5 mL DCM, 0.5 mL TFAadded into the solution at room temperature. Reaction mixture wasstirred at room temperature for 6 hours. The solvent and excess TFA wasremoved by rotary evaporation and dried under vacuum. The crude TFA saltwas dissolved in DCM again and directly submitted for next step.

B (b). 1.1 eq Et₃N was added to free the TFA/amine salt using DCM assolvent. The crude product was purified by column chromatography (100%EtOAc, R_(f). 0.14) to give the product as a white solid (0.1 mmolscale, 24 mg, 65% yield for two steps). ¹H NMR (400 MHz, Methanol-d₄) δ7.55-7.45 (m, 2H), 7.34-7.26 (m, 2H), 4.37 (dtd, J=13.6, 4.0, 1.8 Hz,1H), 3.90 (dtd, J=14.1, 4.0, 1.8 Hz, 1H), 3.82 (tt, J=10.5, 4.1 Hz, 1H),3.22 (ddd, J=14.2, 11.5, 2.9 Hz, 1H), 2.87 (ddd, J=14.0, 11.6, 3.1 Hz,1H), 2.41 (q, J=7.5 Hz, 2H), 2.08-1.87 (m, 2H), 1.47-1.27 (m, 2H), 1.10(t, J=7.5 Hz, 3H); ¹⁹F NMR (376 MHz, Methanol-d₄) δ 35.02; ESI-MS (m/z):374 [MH].

C. Synthesis of 4-(((1s,4s)-4-(3-(4-((fluorosulfonyl)oxy)phenyl) ureido)cyclohexyl)oxy)benzoic Acid

24 mg (0.1 mmol) 4-(((1r,4r)-4-aminocyclohexyl)oxy)benzoic acid wasdissolved in 0.5 mL DMF, 15 μL Et₃N was added to the solution. Reactionmixture stirred at room temperature for 5 min. 0.1 mL 4-isocyanatophenylsulfurofluoridate/DCM solution (0.1 mmol, 1 eq) was added by syringe,followed by 1 mg DABCO. Reaction was heated at 60° C. by an oil bath for24 hours and was monitored by LCMS. Solvent was removed by rotaryevaporation. Crude mixture was directly loaded to a column; purificationby column chromatography (R_(f)=0.41, 100% EtOAc) to give the product asa white solid (0.1 mmol scale, 25 mg, 53% yield for two steps). ¹H NMR(400 MHz, Methanol-d₄) δ 7.99-7.90 (m, 2H), 7.56-7.47 (m, 2H), 7.36-7.26(m, 2H), 7.03-6.92 (m, 2H), 4.43 (ddd, J=10.0, 6.0, 4.0 Hz, 1H),3.73-3.57 (m, 1H), 2.22-2.13 (m, 2H), 2.08 (dt, J=13.6, 3.8 Hz, 2H),1.60 (tdd, J=12.8, 9.9, 3.2 Hz, 2H), 1.45 (tdd, J=13.0, 10.5, 3.2 Hz,2H); ¹⁹F NMR (376 MHz, Methanol-d₄) δ 34.97; ESI-MS (m/z): 453 [MH]⁺.

D. Synthesis of tert-butyl 4-(3-(4-((fluorosulfonyl)oxy)phenyl)ureido)piperidine-1-carboxylate

D (a). 4-Aminophenyl fluorosulfonate was readily prepared and isolatedas a brown solid (mp 41-42° C.) in 91% yield (8.0 g) by reaction of4-aminophenol with sulfuryl fluoride for 6 hours in DCM using 3 eq ofEt₃N. T HNMR (400 MHz, CDCl₃) δ 7.08 (d, J=8.5 Hz, 1H), 6.65 (d, J=8.7Hz, 1H), 3.87 (br s, 1H); ¹³C NMR (101 MHz, CDCl₃) δ 146.9, 142.1,121.8, 115.6, 115.5; ¹⁹F NMR (376 MHz. CDCl₃) δ +35.5; EI-MS (m/z): 191[M]⁺.

D (b). 4-isocyanatophenyl sulfurofluoridate was prepared by reaction of4-Aminophenyl fluorosulfonate with triphosgene and triethylamine ndichloromethane (DCM).

D (c). tert-butyl4-(3-(4-((fluorosulfonyl)oxy)phenyl)ureido)piperidine-1-carboxylate wasprepared from isocyanatophenyl sulfurofluoridate by reaction with theBoc-protected aminopiperadine as shown above and isolated as a whitesolid. The crude product was purified by column chromatography (50:50hexane:EtOAc, R_(f): 0.15) to give the product as a white solid (0.25mmol scale, 93 mg, 88% yield for two steps); ¹H NMR (400 MHz,Methanol-d₄) δ 7.57-7.47 (m, 2H), 7.39-7.23 (m, 2H), 3.99 (dt, J=14.1,3.7 Hz, 2H), 3.75 (tt, J=10.5, 4.1 Hz, 1H), 2.96 (s, 2H), 2.01-1.78 (m,2H), 1.46 (s, 9H), 1.44-1.27 (m, 2H); ¹⁹F NMR (376 MHz, Methanol-d₄) δ35.02; ESI-MS (m/z): 318 [MH]⁺-100.

E. Synthesis of 4-(3-(1-(methylsulfonyl)piperidin-4-yl) ureido)phenylSulfurofluoridate

4-(3-(1-(methylsulfonyl)piperidin-4-yl)ureido)phenyl sulfurofluoridatewas prepared by the same general process as in D(c) above bysubstituting the N-methylsulfonyl piperadine compound for theBoc-protected piperadine compound, as shown above; mp 227-229; ¹H NMR(400 MHz, Methanol-d₄) δ 7.56-7.48 (m, 2H), 7.36-7.27 (m, 2H), 3.76-3.60(m, 3H), 2.96-2.86 (m, 2H), 2.84 (s, 3H), 2.09-1.94 (m, 2H), 1.54 (dtd,J=12.8, 11.0, 4.1 Hz, 2H); ¹⁹F NMR (376 MHz, Methanol-d₄) δ 35.00;ESI-MS (m/z): 396 [MH]⁺.

F. Synthesis of 4-(3-(1-(fluorosulfonyl)piperidin-4-yl)ureido)phenylSulfurofluoridate

F(a). General Procedure for Synthesis of sulfamoylfluorides. Secondaryamine (1 eq), N,N-dimethylaminopyridine (DMAP; 0.5-1 eq) andtriethylamine (2 eq) are mixed in CH₂Cl₂ (0.33 M) in a round-bottomflask filled to one-third capacity. The flask is sealed with septa andevacuated. SO₂F₂ gas is introduced from a balloon. The reaction mixtureis stirred vigorously at room temperature for about 3 to 18 h, andreaction progress is monitored by GC or LC-MS. Upon completion, themixture is concentrated, dissolved in EtOAc, washed with 1 N HCl andbrine, dried over MgSO₄, and concentrated to provide the desiredcompound, usually in quite pure form. In some cases, additionalpurification is performed by passage through a short silica gel column.

F(b). 4-((3-Methylbutylidene)amino)piperidine-1-sulfamoyl fluoride wasprepared according the general procedure F (a), above, with 0.5 eq DMAPand 2 eq Et₃N. Upon completion, the reaction mixture was washed withwater, dried, and concentrated. The product was obtained as a yellow oilyield (1.8 g, 70% yield, accounting for DMAP contamination in ¹H NMR).For characterization, the imine group was hydrolyzed (treatment withisopropanol/water mixture at 50° C. for 1.5 hours) followed by treatmentwith methanesulfonic acid (MsOH) to make the shelf stable salt. EI-MS(m/z): 183 [MH]⁺.

F(c). 4-(3-(1-(fluorosulfonyl)piperidin-4-yl)ureido)phenylsulfurofluoridate

4-(3-(1-(fluorosulfonyl)piperidin-4-yl)ureido)phenyl sulfurofluoridatewas prepared and isolated as a white solid. The crude product waspurified by column chromatography (50:50 hexane:EtOAc, Rf: 0.57) to givethe product as a white solid; mp 168-170° C.; 1H NMR (400 MHz,Methanol-d₄) δ 7.54-7.48 (m, 2H), 7.33-7.27 (m, 2H), 3.88-3.80 (m, 2H),3.77 (td, J=6.4, 3.2 Hz, 1H), 3.24 (ddt, J=13.1, 11.5, 3.0 Hz, 2H),2.11-1.98 (m, 2H), 1.59 (dtd, J=13.7, 11.0, 4.2 Hz, 2H); ¹⁹F NMR (376MHz, Methanol-d₄) δ 35.06; ESI-MS (m/z): 400 [MH]⁺.

G. Synthesis of 4-(3-(4-ethynylphenyl)ureido)piperidine-1-sulfonylFluoride

4-(3-(4-ethynylphenyl)ureido)piperidine-1-sulfonylfluoride was preparedand isolated as a white solid according to the general procedure in F(a)above from 1-ethynyl-4-isocyanatobenzene. The crude product was purifiedby column chromatography (50:50 hexane:EtOAc, R_(f): 0.70) to give theproduct as a white solid; mp 227-229° C.; ¹H NMR (400 MHz, Methanol-d₄)δ 7.33 (s, 4H), 3.89-3.80 (m, 2H), 3.80-3.70 (m, 1H), 3.34 (s, 1H),3.27-3.19 (m, 2H), 2.11-1.97 (m, 2H), 1.68-1.52 (m, 2H); ¹⁹F NMR (376MHz, Methanol-d₄) δ 39.06; ESI-MS (m/z): 326 [MH]⁺.

H. Synthesis of4-(3-(4-(fluorosulfonyl)phenyl)ureido)piperidine-1-sulfonyl Fluoride

4-(3-(4-(fluorosulfonyl)phenyl)ureido)piperidine-1-sulfonyl fluoride wasisolated as a white solid according to the general procedure in F(a)above from 4-isocyanatobenzenesulfonyl fluoride. The crude product waspurified by column chromatography (50:50 hexane:EtOAc; R_(f): 0.56) togive the product as a yellow gel; ¹H NMR (400 MHz, Methanol-d₄) δ7.93-7.86 (m, 2H), 7.72-7.65 (m, 2H), 3.92-3.73 (m, 3H), 3.29-3.21 (m,2H), 2.06 (dt, J=13.2, 3.8 Hz, 2H), 1.71-1.52 (m, 2H); ¹⁹F NMR (376 MHz,Methanol-d₄) δ 65.58, 39.15; ESI-MS (m/z): 384 [MH]⁺.

I. Synthesis of2-(4-(3-(4-(trifluoromethoxy)phenyl)ureido)piperidin-1-yl)ethane-1-sulfonylFluoride

I(a). General procedure for the reaction of primary and secondary amineswith ESF (adapted from Krutak, J. J.; Burpitt, R. D.; Moore, W. H.;Hyatt, J. A. J. Org. Chem. 1979, 44, 3847-3858). The starting amine (1equiv) is dissolved in organic solvent (usually CH₂Cl₂ or THF, 0.1-0.5 Min substrate) and treated with ESF (1-2.5 equiv). The reaction mixtureis stirred at room temperature for several minutes to several hours,monitoring conversion by LC-MS. Upon completion, the solvent and excessof ESF are removed by rotary evaporation and dried under vacuum, usuallyproviding clean product. When purification by column chromatography ismentioned, it was done to remove trace impurities.

I (b).2-(4-(3-(4-(trifluoromethoxy)phenyl)ureido)piperidin-1-yl)ethane-1-sulfonylfluoride was prepared and isolated as a yellow solid (quantitativeyield. 22 mg) according to the general ESF procedure I(a), above. Thepiperidine compound was mixed with 1.1 eq ESF (ethenesulfonyl fluoride)in DCM/CH₃CN at room temperature. Reaction finished in 10 minutes. mp180-182° C.; ¹H NMR (400 MHz, Methanol-d₄) δ 7.54-7.39 (m, 2H),7.23-7.01 (m, 2H), 4.37 (d, J=13.1 Hz, 2H), 3.85 (s, 1H), 3.77-3.48 (m,4H), 3.14 (s, 2H), 2.21-2.07 (m, 2H), 1.92 (d, J=12.8 Hz, 2H); ¹⁹F NMR(376 MHz, Methanol-d₄) δ 54.78, −59.37; ESI-MS (m/z): 414 [MH]⁺.

The fluorosulfate, ESF, and fluorosulfamate derivatives described inthis example were tested in an assay for sEH inhibition activity. Thecompounds exhibited IC₅₀ values against human sEH of less than 10 nM,with some having IC₅₀ values of less than 1 nM.

Example 11. Naproxen-SF Derivative

Naproxen was extracted from commercially available caplet and wastreated with 48% aqueous solution of hydrobromic acid under refluxcondition to remove the methyl group from the methoxy substituent. Aftercompletion of the reaction and cooling the mixture to room temperature,the demethylated naproxen was obtained as yellow needle-like crystals.The crystals were suspended in a solvent comprising dichloromethane andwater (3:2 v/v). Triethylamine (2 equiv) was added to the suspendedcrystals and the resulting mixture was stirred under a nitrogenatmosphere for about 10 minutes, followed by an atmosphere of sulfurylfluoride (supplied by a balloon filled with sulfuryl fluoride sealed tothe reaction vessel). After completion of the reaction of the sulfurylfluoride with the demethylated hydroxyl group of the naproxen, volatileswere removed under reduced pressure. A solution of 1M hydrochloric acidwas used to adjust the pH of aqueous phase to neutral or weak acidic,then the aqueous phase was extracted with ethyl acetate. The organicphase was then isolated, washed with brine, and dried over anhydroussodium sulfate. After filtration and concentration, the naproxen-SFproduct was purified and isolated by flash column chromatography.

1H NMR (400 MHz, d6-DMSO) 12.45 (s, 1H), 8.20 (d, J=2.8 Hz, 1H), 8.15(d, J=9.2 Hz, 1H), 8.04 (d, J=8.8 Hz, 1H), 7.96 (s, 1H), 7.69 (dd,J=9.2, and 2.8 Hz, 1H), 7.62 (dd, J=8.2, and 2.0 Hz, 1H), 3.91 (q,J=7.2, 1H), 1.47 (d, J=6.8, 3H); 13C NMR (101 MHz, d6-DMSO); 175.0,147.1, 140.7, 132.3, 131.9, 130.9, 128.3, 128.0, 126.0, 119.4, 118.7,44.7, 18.3; 19F NMR (376 MHz, d6-DMSO) 39.03; melting point: 146-147° C.(hexane/ethyl acetate).

Example 12. Paracetamol-SF Derivative

Paracetamol was suspended in dichloromethane under a nitrogen nitrogenatmosphere and triethylamine (1.5 equiv) was added. The mixture wasstirred for 10 minutes, and then a sulfuryl fluoride was introduced (viaa balloon filled with sulfuryl fluoride) to form the paracetamol-SFanalog. After completion of the reaction, the solvent was removed underreduced pressure, and the residue was dissolved in ethyl acetate.Filtration removed insoluble salts and the solution was concentrated invacuo. The crude product was purified by flash column chromatography.

1H NMR (400 MHz, CDCl3) 7.63 (d, J=8.8 Hz, 2H), 7.30 (d, J=8.8 Hz, 2H),2.20 (s, 3H); 13C NMR (101 MHz, CDCl3) 168.6, 145.9, 138.3, 121.7,121.3, 24.7; 19F NMR (376 MHz, CDCl3) 36.91; melting point: 152-153° C.

All references, including publications, patent applications, andpatents, cited herein are hereby incorporated by reference to the sameextent as if each reference were individually and specifically indicatedto be incorporated by reference and were set forth in its entiretyherein.

Preferred embodiments of this invention are described herein, includingthe best mode known to the inventors for carrying out the invention.Variations of those preferred embodiments may become apparent to thoseof ordinary skill in the art upon reading the foregoing description. Theinventors expect skilled artisans to employ such variations asappropriate, and the inventors intend for the invention to be practicedotherwise than as specifically described herein. Accordingly, thisinvention includes all modifications and equivalents of the subjectmatter recited in the claims appended hereto as permitted by applicablelaw. Moreover, any combination of the above-described elements in allpossible variations thereof is encompassed by the invention unlessotherwise indicated herein or otherwise clearly contradicted by context.

We claim:
 1. A modified amino acid comprising an amino acid core groupbound to a SO₂F or CH₂CH₂SO₂F group; wherein the amino acid core grouphas a side chain comprising a hydroxyl or amino substituent, and theSO₂F or CH₂CH₂SO₂F group is bound to the side chain in place of ahydrogen of the hydroxyl or amino substituent thereof.
 2. The modifiedamino acid of claim 1, wherein the amino acid core group is tyrosine. 3.The modified amino acid of claim 1, wherein the amino acid core group is2,6-dimethyltyrosine.
 4. The modified amino acid of claim 1, wherein theamino acid core group is histidine.
 5. The modified amino acid of claim1, wherein the amino acid core group is lysine.
 6. The modified aminoacid of claim 1, wherein the amino acid core group is arginine.
 7. Apolypeptide comprising a modified amino acid residue bearing a SO₂F orCH₂CH₂SO₂F group; wherein the a modified amino acid residue comprises anamino acid residue core group having a side chain comprising a hydroxylor amino substituent, and the SO₂F or CH₂CH₂SO₂F group is bound to theside chain in place of a hydrogen of the hydroxyl or amino substituentthereof.
 8. The polypeptide of claim 7, wherein the amino acid residuecore group is a tyrosine residue.
 9. The polypeptide of claim 7, whereinthe amino acid residue core group is a 2,6-dimethyltyrosine residue. 10.The polypeptide of claim 7, wherein the amino acid residue core group isa histidine residue.
 11. The polypeptide of claim 7, wherein the aminoacid residue core group is a lysine residue.
 12. The polypeptide ofclaim 7, wherein the amino acid residue core group is an arginineresidue.
 13. The polypeptide of claim 7, wherein the polypeptide is aprotein.
 14. A method for covalently attaching a SO₂F or CH₂CH₂SO₂Fsubstituted compound to a receptor protein; the method comprisingcontacting the compound with the receptor protein in vivo; wherein: thereceptor protein comprises a receptor site that includes a nucleophilicamino acid side chain in the receptor site; the compound is apolypeptide comprising a modified amino acid residue bearing a SO₂F orCH₂CH₂SO₂F group; wherein the modified amino acid residue comprises anamino acid residue core group having a side chain comprising a hydroxylor amino substituent, and the SO₂F or CH₂CH₂SO₂F group is bound to theside chain in place of a hydrogen of the hydroxyl or amino substituentthereof, the compound has a molecular configuration that can interactwith and dock with the receptor site; and when docked with the receptorsite, the SO₂F or CH₂CH₂SO₂F group of the compound is oriented withrespect to the nucleophilic side chain of the receptor site such thatthe nucleophilic side chain of the receptor site displaces F from theSO₂F or CH₂CH₂SO₂F group and forms a covalent bond to the S atomthereof.
 15. The method of claim 14, wherein the amino acid residue coregroup is a tyrosine residue.
 16. The method of claim 14, wherein theamino acid residue core group is a 2,6-dimethyltyrosin residue.
 17. Themethod of claim 14, wherein the amino acid residue core group is ahistidine residue.
 18. The method of claim 14, wherein the amino acidresidue core group is a lysine residue.
 19. The method of claim 14,wherein the amino acid residue core group is an arginine residue. 20.The method of claim 14, wherein the polypeptide is a protein.